Compositions and methods for treating leber&#39;s hereditary optic neuropathy

ABSTRACT

Disclosed herein is a recombinant nucleic acid, comprising: a mitochondrial targeting sequence; a mitochondrial protein coding sequence, wherein said mitochondrial protein coding sequence encodes a polypeptide comprising a mitochondrial protein; and a 3′UTR nucleic acid sequence. Also disclosed is a pharmaceutical composition comprising the recombinant nucleic acid and a method of treating Leber&#39;s hereditary optic neuropathy (LHON) using the pharmaceutical composition.

CROSS-REFERENCE

This application claims the benefit of PCT Application No.PCT/CN2018/095023, filed on Jul. 9, 2018; PCT Application No.PCT/CN2018/103937, filed on Sep. 4, 2018; Chinese Application Nos.CN201810703168.7 and CN201810702492.7, both filed on Jun. 29, 2018; PCTApplication No. PCT/CN2018/113799, filed on Nov. 2, 2018; ChineseApplication No. CN201811230856.2, filed on Oct. 22, 2018; PCTApplication No. PCT/CN2018/118662, filed on Nov. 30, 2018; ChineseApplication No. CN201811221305.X, filed on Oct. 19, 2018; PCTApplication No. PCT/CN2019/070461, filed on Jan. 4, 2019; ChineseApplication No. CN201810948193.1, filed on Aug. 20, 2018; all of whichare incorporated herein by reference in their entirety.

REFERENCE TO A SEQUENCE LISTING

The instant application contains a Sequence Listing which has beensubmitted in ASCII format via EFS-Web and is hereby incorporated byreference in its entirety. Said ASCII copy, created on Jun. 30, 2019, isnamed 207298476_1.txt and is 304,914 bytes in size.

BACKGROUND OF THE INVENTION

Lebers hereditary optic neuropathy (LHON) is a mitochondrially inherited(transmitted from mother to offspring) degeneration of retinal ganglioncells (RGCs) and their axons that leads to an acute or subacute loss ofcentral vision; this affects predominantly young adult males. LHON isonly transmitted through the mother, as it is primarily due to mutationsin the mitochondrial (not nuclear) genome, and only the egg contributesmitochondria to the embryo. LHON is usually due to one of threepathogenic mitochondrial DNA (mtDNA) point mutations. These mutationsare at nucleotide positions 11778 G to A (G11778A), 3460 G to A (G3460A)and 14484 T to C (TI 4484C), respectively in the NADH dehydrogenasesubunit-4 protein (ND4), NADH dehydrogenase subunit-1 protein (ND1) andNADH dehydrogenase subunit-6 protein (ND6) subunit genes of complex I ofthe oxidative phosphorylation chain in mitochondria. Each mutation isbelieved to have significant risk of permanent loss of vision. Ittypically progresses within several weeks to several months withoutpain, until the binocular vision deteriorate to below 0.1, whichseriously affects the quality of life of the patient. Two LHON mutants,G3460A and T14484C, results in the reduction of the patient's plateletsisolated mitochondrial NADH dehydrogenase activity by 80%. Ninetypercent of the Chinese LHON patients carry the G11778A mutation. TheG11778A mutation changes an arginine into histidine in the ND4 protein,resulting the dysfunction and optic nerve damage in LHON patients. Thereis a need for developing compositions and methods for treating LHON withhigher transfection efficiency and treatment efficacy.

SUMMARY OF THE INVENTION

Disclosed here recombinant nucleic acids, pharmaceutical compositions,and methods for treating LHON. In one aspect, disclosed herein is arecombinant nucleic acid, comprising: a mitochondrial targetingsequence; a mitochondrial protein coding sequence comprising a sequencethat is at least 99% identical to a sequence selected from the groupconsisting of SEQ ID NO: 7, 8, 10, and 12; and a 3′UTR nucleic acidsequence.

In some cases, the mitochondrial targeting sequence encodes apolypeptide comprising a peptide sequence that is at least 90%, at least95%, at least 97%, at least 99%, or 100% identical to a sequenceselected from the group consisting of SEQ ID NO: 129-159. In some cases,the mitochondrial targeting sequence comprises a sequence that is atleast 90%, at least 95%, at least 97%, at least 99%, or 100% identicalto a sequence as set forth in SEQ ID NO: 2. In some cases, themitochondrial targeting sequence comprises a sequence that is at least90%, at least 95%, at least 97%, at least 99%, or 100% identical to asequence as set forth in SEQ ID NO: 3. In some cases, the mitochondrialtargeting sequence comprises a sequence that is at least 90%, at least95%, at least 97%, at least 99%, or 100% identical to a sequence as setforth in SEQ ID NO: 4. In some cases, the mitochondrial targetingsequence comprises a sequence that is at least 90%, at least 95%, atleast 97%, at least 99%, or 100% identical to a sequence as set forth inSEQ ID NO: 5.

In some cases, the mitochondrial protein coding sequence comprises asequence that is at least 90%, at least 95%, at least 97%, at least 99%,or 100% identical to a sequence as set forth in SEQ ID NO: 7 or 8. Insome cases, the mitochondrial protein coding sequence comprises asequence that is at least 90%, at least 95%, at least 97%, at least 99%,or 100% identical to a sequence as set forth in SEQ ID NO: 10. In somecases, the mitochondrial protein coding sequence comprises a sequencethat is at least 90%, at least 95%, at least 97%, at least 99%, or 100%identical to a sequence as set forth in SEQ ID NO: 12.

In some cases, the 3′UTR nucleic acid sequence comprises a sequence thatis at least 90%, at least 95%, at least 97%, at least 99%, or 100%identical to a sequence selected from the group consisting of SEQ ID NO:111-125. In some cases, the 3′UTR nucleic acid sequence comprises asequence that is at least 90%, at least 95%, at least 97%, at least 99%,or 100% identical to a sequence as set forth in SEQ ID NO: 13 or SEQ IDNO: 14.

In some cases, the recombinant nucleic acid comprises a sequence that isat least 90%, at least 95%, at least 97%, at least 99%, or 100%identical to a sequence selected from the group consisting of SEQ ID NO:17-20, 23-24, 27-28, 31-34, 37-38, 41-42, 45-48, 51-52, 55-56, 59-62,65-66, 69-70, 73-76, 79-80, and 83-84.

In another aspect, disclosed herein is a recombinant nucleic acid,comprising: a mitochondrial targeting sequence comprising a sequencethat is at least 90% identical to a sequence selected from the groupconsisting of SEQ ID NO: 2, 3, 4, and 5; a mitochondrial protein codingsequence, wherein the mitochondrial protein coding sequence encodes apolypeptide comprising a mitochondrial protein; and a 3′UTR nucleic acidsequence.

In some cases, the mitochondrial targeting sequence comprises a sequencethat is at least 90%, at least 95%, at least 97%, at least 99%, or 100%identical to a sequence as set forth in SEQ ID NO: 2. In some cases, themitochondrial targeting sequence comprises a sequence that is at least90%, at least 95%, at least 97%, at least 99%, or 100% identical to asequence as set forth in SEQ ID NO: 3. In some cases, the mitochondrialtargeting sequence comprises a sequence that is at least 90%, at least95%, at least 97%, at least 99%, or 100% identical to a sequence as setforth in SEQ ID NO: 4. In some cases, the mitochondrial targetingsequence comprises a sequence that is at least 90%, at least 95%, atleast 97%, at least 99%, or 100% identical to a sequence as set forth inSEQ ID NO: 5.

In some cases, the mitochondrial protein is selected from a groupconsisting of NADH dehydrogenase 4 (ND4), NADH dehydrogenase 6 (ND6),NADH dehydrogenase 1 (ND1), and a variant thereof. In some cases, themitochondrial protein comprises NADH dehydrogenase 4 (ND4), or a variantthereof. In some cases, the mitochondrial protein comprises a peptidesequence that is at least 90%, at least 95%, at least 97%, at least 99%,or 100% identical to a sequence as set forth in SEQ ID NO: 160. In somecases, the mitochondrial protein coding sequence comprises a sequencethat is at least 90%, at least 95%, at least 97%, at least 99%, or 100%identical to a sequence as set forth in SEQ ID NO: 6, 7, or 8. In somecases, the mitochondrial protein comprises NADH dehydrogenase 6 (ND6),or a variant thereof. In some cases, the mitochondrial protein comprisesa sequence that is at least 90%, at least 95%, at least 97%, at least99%, or 100% identical to a sequence as set forth in SEQ ID NO: 161. Insome cases, the mitochondrial protein coding sequence comprises asequence that is at least 90%, at least 95%, at least 97%, at least 99%,or 100% identical to a sequence as set forth in SEQ ID NO: 9 or 10. Insome cases, the mitochondrial protein comprises NADH dehydrogenase 1(ND1), or a variant thereof. In some cases, the mitochondrial proteincomprises a sequence that is at least 90/o, at least 95%, at least 97%,at least 99%, or 100% identical to a sequence as set forth in SEQ ID NO:162. In some cases, the mitochondrial protein coding sequence comprisesa sequence that is at least 90%, at least 95%, at least 97%, at least99%, or 100% identical to a sequence as set forth in SEQ ID NO: 11 or12.

In some cases, the 3′UTR nucleic acid sequence is located at 3′ of themitochondrial targeting sequence. In some cases, the 3′UTR nucleic acidsequence comprises a sequence selected from the group consisting ofhsACO2, hsATP5B, hsAK2, hsALDH2, hsCOX10, hsUQCRFS1, hsNDUFV1, hsNDUFV2,hsSOD2, hsCOX6c, hsIRPl, hsMRPS12, hsATP5J2, mSOD2, and hsOXA1L. In somecases, the 3′UTR nucleic acid sequence comprises a sequence that is atleast 90%, at least 95%, at least 97%, at least 99%, or 100% identicalto a sequence selected from the group consisting of SEQ ID NO: 111-125.In some cases, the 3′UTR nucleic acid sequence comprises a sequence thatis at least 90%, at least 95%, at least 97%, at least 99%, or 100%identical to a sequence as set forth in SEQ ID NO: 13 or SEQ ID NO: 14.

In some cases, the mitochondrial targeting sequence is located at 5′ ofthe 3′UTR nucleic acid sequence. In some cases, the mitochondrialtargeting sequence is located at 3′ of the mitochondrial targetingsequence.

In some cases, the recombinant nucleic acid comprises a sequence that isat least 90%, at least 95%, at least 97%, at least 99%, or 100%identical to a sequence selected from the group consisting of SEQ ID NO:29-84.

In another aspect, disclosed herein is a recombinant nucleic acid,comprising: a mitochondrial targeting sequence; a mitochondrial proteincoding sequence comprising a sequence that is at least 90%, at least95%, at least 97%, at least 99%, or 100% identical to a sequenceselected from the group consisting of SEQ ID NO: 7, 8, 10, and 12; and a3′UTR nucleic acid sequence.

In some cases, the mitochondrial targeting sequence comprises a sequenceencodes a polypeptide selected from the group consisting of hsCOX10,hsCOX8, scRPM2, lcSirt5, tbNDUS7, ncQCR2, hsATP5G2, hsLACTB, spilv1,gmCOX2, crATP6, hsOPA1, hsSDHD, hsADCK3, osP0644B06.24-2, Neurosporacrassa ATP9 (ncATP9), hsGHITM, hsNDUFAB1, hsATP5G3, crATP6_hsADCK3,ncATP9_ncATP9, zmLOC100282174, ncATP9_zmLOC100282174_spilv1_ncATP9,zmLOC100282174_hsADCK3_crATP6_hsATP5G3, zmLOCIO0282174_hsADCK3_hsATP5G3,ncATP9_zmLOC100282174, hsADCK3 zmLOC100282174 crATP6 hsATP5G3,crATP6_hsADCK3_zmLOC100282174_hsATP5G3, hsADCK3_zmLOC100282174,hsADCK3_zmLOC100282174_crATP6, ncATP9_zmLOC100282174_spilv1_GNFP_ncATP9,and ncATP9_zmLOC100282174_spilv1_cSirtS_osP0644B06.24-2_hsATP5G2_ncATP9.In some cases, the mitochondrial targeting sequence encodes apolypeptide comprising a peptide sequence that is at least 90%, at least95%, at least 97%, at least 99%, or 100% identical to a sequenceselected from the group consisting of SEQ ID NO: 129-159. In some cases,the mitochondrial targeting sequence comprises a sequence that is atleast 90%, at least 95%, at least 97%, at least 99%, or 100% identicalto a sequence as set forth in SEQ ID NO: 2 or 3. In some cases, themitochondrial targeting sequence comprises a sequence that is at least90%, at least 95%, at least 97%, at least 99%, or 100% identical to asequence as set forth in SEQ ID NO: 4. In some cases, the mitochondrialtargeting sequence comprises a sequence that is at least 90%, at least95%, at least 97%, at least 99%, or 100% identical to a sequence as setforth in SEQ ID NO: 5.

In some cases, the mitochondrial protein coding sequence comprises asequence that is at least 90%, at least 95%, at least 97%, at least 99%,or 100% identical to a sequence as set forth in SEQ ID NO: 7 or 8. Insome cases, the mitochondrial protein coding sequence comprises asequence that is at least 90%, at least 95%, at least 97%, at least 99%,or 100% identical to a sequence as set forth in SEQ ID NO: 10. In somecases, the mitochondrial protein coding sequence comprises a sequencethat is at least 90%, at least 95%, at least 97%, at least 99%, or 100%identical to a sequence as set forth in SEQ ID NO: 12.

In some cases, the 3′UTR nucleic acid sequence is located at 3′ of themitochondrial targeting sequence. In some cases, the 3′UTR nucleic acidsequence comprises a sequence selected from the group consisting ofhsACO2, hsATPSB, hsAK2, hsALDH2, hsCOX10, hsUQCRFS1, hsNDUFV1, hsNDUFV2,hsSOD2, hsCOX6c, hs1RP1, hsMRPS12, hsATP5J2, mSOD2, and hsOXA1L. In somecases, the 3′UTR nucleic acid sequence comprises a sequence that is atleast 90%, at least 95%, at least 97%, at least 99%, or 100% identicalto a sequence selected from the group consisting of SEQ ID NO: 111-125.In some cases, the 3′UTR nucleic acid sequence comprises a sequence thatis at least 90%, at least 95%, at least 97%, at least 99%, or 100%identical to a sequence as set forth in SEQ ID NO: 13 or SEQ ID NO: 14.

In some cases, the mitochondrial targeting sequence is located at 5′ ofthe 3′UTR nucleic acid sequence. In some cases, the mitochondrialtargeting sequence is located at 3′ of the mitochondrial targetingsequence.

In some cases, the recombinant nucleic acid comprises a sequence that isat least 90%, at least 95%, at least 97%, at least 99%, or 100%identical to a sequence selected from the group consisting of SEQ ID NO:17-20, 23-24, 27-28, 31-34, 37-38, 41-42, 4548, 51-52, 55-56, 59-62,65-66, 69-70, 73-76, 79-80, and 83-84.

In another aspect, disclosed herein is a recombinant nucleic acid,comprising a mitochondrial targeting sequence that is at least 90%, atleast 95%, at least 97%, at least 99%, or 100% identical to a sequenceselected from the group consisting of SEQ ID NO: 2, 3, and 4. In somecases, the mitochondrial targeting sequence comprises a sequence that isat least 90%, at least 95%, at least 97%, at least 99%, or 100%identical to a sequence as set forth in SEQ ID NO: 2. In some cases, themitochondrial targeting sequence comprises a sequence that is at least90%, at least 95%, at least 97%, at least 99%, or 100% identical to asequence as set forth in SEQ ID NO: 3. In some cases, the mitochondrialtargeting sequence comprises a sequence that is at least 90%, at least95%, at least 97%, at least 99%, or 100% identical to a sequence as setforth in SEQ ID NO: 4.

In some cases, the recombinant nucleic acid further comprises amitochondrial protein coding sequence, wherein the mitochondrial proteincoding sequence encodes a polypeptide comprising a mitochondrialprotein. In some cases, the mitochondrial protein is selected from agroup consisting of NADH dehydrogenase 4 (ND4), NADH dehydrogenase 6(ND6), NADH dehydrogenase 1 (ND1), and a variant thereof. In some cases,the mitochondrial protein comprises NADH dehydrogenase 4 (ND4), or avariant thereof. In some cases, the mitochondrial protein comprises apeptide sequence that is at least 90%, at least 95%, at least 97%, atleast 99%, or 100% identical to a sequence as set forth in SEQ ID NO:160. In some cases, the mitochondrial protein coding sequence comprisesa sequence that is at least 90%, at least 95%, at least 97%, at least99%, or 100% identical to a sequence as set forth in SEQ ID NO: 6, 7, or8. In some cases, the mitochondrial protein comprises NADH dehydrogenase6 (ND6), or a variant thereof. In some cases, the mitochondrial proteincomprises a sequence that is at least 90%, at least 95%, at least 97%,at least 99%, or 100% identical to a sequence as set forth in SEQ ID NO:161. In some cases, the mitochondrial protein coding sequence comprisesa sequence that is at least 90%, at least 95%, at least 97%, at least99%, or 100% identical to a sequence as set forth in SEQ ID NO: 9 or 10.In some cases, the mitochondrial protein comprises NADH dehydrogenase 1(ND1), or a variant thereof. In some cases, the mitochondrial proteincomprises a sequence that is at least 90%, at least 95%, at least 97%,at least 99%, or 100% identical to a sequence as set forth in SEQ ID NO:162. In some cases, the mitochondrial protein coding sequence comprisesa sequence that is at least 90%, at least 95%, at least 97%, at least99%, or 100% identical to a sequence as set forth in SEQ ID NO: 11 or12.

In some cases, the recombinant nucleic acid further comprises a 3′UTRnucleic acid sequence. In some cases, the 3′UTR nucleic acid sequence islocated at 3′ of the mitochondrial targeting sequence. In some cases,the 3′UTR nucleic acid sequence comprises a sequence selected from thegroup consisting of hsACO2, hsATP5B, hsAK2, hsALDH2, hsCOX10, hsUQCRFS1,hsNDUFV1, hsNDUFV2, hsSOD2, hsCOX6c, hsIRPl, hsMRPS12, hsATP5J2, mSOD2,and hsOXA1L. In some cases, the 3′UTR nucleic acid sequence comprises asequence that is at least 90%, at least 95%, at least 97%, at least 99%,or 100% identical to a sequence selected from the group consisting ofSEQ ID NO: 111-125. In some cases, the 3′UTR nucleic acid sequencecomprises a sequence that is at least 90%, at least 95%, at least 97%,at least 99%, or 100% identical to a sequence as set forth in SEQ ID NO:13 or SEQ ID NO: 14. In some cases, the mitochondrial targeting sequenceis located at 5′ of the 3′UTR nucleic acid sequence. In some cases, themitochondrial targeting sequence is located at 3′ of the mitochondrialtargeting sequence.

In some cases, the recombinant nucleic acid comprises a sequence that isat least 90%, at least 95%, at least 97%, at least 99%, or 100%identical to a sequence selected from the group consisting of SEQ ID NO:29-70.

In another aspect, disclosed herein is a recombinant nucleic acid,comprising a mitochondrial protein coding sequence, wherein themitochondrial protein coding sequence encodes a polypeptide comprising amitochondrial protein, wherein the mitochondrial protein coding sequencecomprises a sequence that is at least 90%, at least 95%, at least 97%,at least 99%, or 100% identical to a sequence selected from the groupconsisting of SEQ ID NO: 7, 8, 10, and 12.

In some cases, the recombinant nucleic acid further comprises amitochondrial targeting sequence. In some cases, the mitochondrialtargeting sequence comprises a sequence encodes a polypeptide selectedfrom the group consisting of hsCOX10, hsCOX8, scRPM2, lcSirt5, tbNDUS7,ncQCR2, hsATP5G2, hsLACTB, spilv1, gmCOX2, crATP6, hsOPA1, hsSDHD,hsADCK3, osP0644B06.24-2, Neurospora crassa ATP9 (ncATP9), hsGHITM,hsNDUFAB1, hsATP5G3, crATP6_hsADCK3, ncATP9_ncATP9, zmLOC100282174,ncATP9_zmLOC100282174_spily i_ncATP9,zmLOC100282174_hsADCK3_crATP6_hsATP5G3, zmLOC100282174_hsADCK3_hsATP5G3,ncATP9_zmLOC100282174, hsADCK3_zmLOC100282174_crATP6_hsATP5G3,crATP6_hsADCK3_zmLOC100282174_hsATP5G3, hsADCK3_zmLOC100282174,hsADCK3_zmLOC100282174_crATP6, ncATP9_zmLOC100282174_spilv1_GNFP_ncATP9,andncATP9_zmLOC100282174_spilv1_lcSirt5_osP0644B06.24-2_hsATP5G2_ncATP9. Insome cases, the mitochondrial targeting sequence encodes a polypeptidecomprising a peptide sequence that is at least 90%, at least 95%, atleast 97%, at least 99%, or 100% identical to a sequence selected fromthe group consisting of SEQ ID NO: 129-159. In some cases, themitochondrial targeting sequence comprises a sequence that is at least90%, at least 95%, at least 97%, at least 99%, or 100% identical to asequence as set forth in SEQ ID NO: 2. In some cases, the mitochondrialtargeting sequence comprises a sequence that is at least 90%, at least95%, at least 97%, at least 99%, or 100% identical to a sequence as setforth in SEQ ID NO: 3. In some cases, the mitochondrial targetingsequence comprises a sequence that is at least 90%, at least 95%, atleast 97%, at least 99%, or 100% identical to a sequence as set forth inSEQ ID NO: 4. In some cases, the mitochondrial targeting sequencecomprises a sequence that is at least 90%, at least 95%, at least 97%,at least 99%, or 100% identical to a sequence as set forth in SEQ ID NO:5.

In some cases, the mitochondrial protein coding sequence comprises asequence that is at least 90%, at least 95%, at least 97%, at least 99%,or 100% identical to a sequence as set forth in SEQ ID NO: 7 or 8. Insome cases, the mitochondrial protein coding sequence comprises asequence that is at least 90%, at least 95%, at least 97%, at least 99%,or 100% identical to a sequence as set forth in SEQ ID NO: 10. In somecases, the mitochondrial protein coding sequence comprises a sequencethat is at least 90%, at least 95%, at least 97/a, at least 99%, or 100%identical to a sequence as set forth in SEQ ID NO: 12.

In some cases, the recombinant nucleic acid further comprises a3′UTRnucleic acid sequence. In some cases, the 3′UTR nucleic acid sequence islocated at 3′ of the mitochondrial targeting sequence. In some cases,the 3′UTR nucleic acid sequence comprises a sequence selected from thegroup consisting of hsACO2, hsATPSB, hsAK2, hsALDH2, hsCOX10, hsUQCRFS1,hsNDUFV1, hsNDUFV2, hsSOD2, hsCOX6c, hslRP1, hsMRPS12, hsATP5J2, mSOD2,and hsOXA1L. In some cases, the 3′UTR nucleic acid sequence comprises asequence that is at least 90%, at least 95%, at least 97%, at least 99%,or 100% identical to a sequence selected from the group consisting ofSEQ ID NO: 111-125. In some cases, the 3′UTR nucleic acid sequencecomprises a sequence that is at least 90%, at least 95%, at least 97%,at least 99%, or 100% identical to a sequence as set forth in SEQ ID NO:13 or SEQ ID NO: 14. In some cases, the mitochondrial targeting sequenceis located at 5′ of the 3′UTR nucleic acid sequence. In some cases, themitochondrial targeting sequence is located at 3′ of the mitochondrialtargeting sequence.

In some cases, the recombinant nucleic acid comprises a sequence that isat least 90%, at least 95%, at least 97%, at least 99%, or 100%identical to a sequence selected from the group consisting of SEQ ID NO:17-20, 23-24, 27-28, 31-34, 37-38, 41-42, 45-48, 51-52, 55-56, 59-62,65-66, 69-70, 73-76, 79-80, and 83-84.

In another aspect, disclosed herein is a viral vector comprising therecombinant nucleic acid disclosed herein. In some cases, the viralvector is an adeno-associated virus (AAV) vector. In some cases, the AAVvector is selected from the group consisting ofAAV1, AAV2, AAV3, AAV4,AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV12, AAV13, AAV14, AAV15,and AAV16 vectors. In some cases, the AAV vector is a recombinant AAV(rAAV) vector. In some cases, the rAAV vector is rAAV2 vector.

In another aspect, disclosed herein is a pharmaceutical composition,comprising an adeno-associated virus (AAV) comprising any recombinantnucleic acid disclosed herein. In some cases, the pharmaceuticalcomposition further comprises a pharmaceutically acceptable excipientthereof. Also disclosed is a pharmaceutical composition, comprising theviral vector disclosed herein, and a pharmaceutically acceptableexcipient thereof, wherein the viral vector comprises any recombinantnucleic acid disclosed herein. Also disclosed is a pharmaceuticalcomposition, comprising: an adeno-associated virus (AAV) comprising anyrecombinant nucleic acid disclosed herein, wherein the recombinantnucleic acid comprises a sequence that is at least 90%, at least 95%, atleast 97%, at least 99%, or 100% identical to a sequence as set forth inSEQ ID NO: 15; and a pharmaceutically acceptable excipient.

In some cases, the pharmaceutically acceptable excipient comprisesphosphate-buffered saline (PBS), α,α-trehalose dehydrate, L-histidinemonohydrochloride monohydrate, polysorbate 20, NaCl, NaH₂PO₄. Na₂HPO₄,KH₂PO₄, K₂HPO₄, poloxamer 188, or any combination thereof. In somecases, the pharmaceutically acceptable excipient is selected fromphosphate-buffered saline (PBS), α,α-trehalose dehydrate, L-histidinemonohydrochloride monohydrate, polysorbate 20, NaCl, NaH₂PO₄, Na₂HPO₄,KH₂PO₄, K₂HPO₄, poloxamer 188, and any combination thereof. In somecases, the pharmaceutically acceptable excipient comprises poloxamer188. In some cases, the pharmaceutically acceptable excipient comprises0.0001%-0.01% poloxamer 188. In some cases, the pharmaceuticallyacceptable excipient comprises 0.001% poloxamer 188. In some cases, thepharmaceutically acceptable excipient further comprises one or moresalts. In some cases, the one or more salts comprises NaCl, NaH₂PO₄,Na₂HPO₄, and KH₂PO₄. In some cases, the one or more salts comprises 80mM NaCl, 5 mM NaH₂PO₄, 40 mM Na₂HPO₄, and 5 mM KH₂PO₄. In some cases,the pharmaceutical composition has a pH of 6-8. In some cases, thepharmaceutical composition has a pH of 7.2-7.4. In some cases, thepharmaceutical composition has a pH of 7.3. In some cases, thepharmaceutical composition has a viral titer of at least 1.0×10¹⁰ vg/mL.In some cases, the pharmaceutical composition has a viral titer of atleast 5.0×10¹⁰ vg/mL.

In some cases, the pharmaceutical composition is subject to fivefreeze/thaw cycles, the pharmaceutical composition retains at least 60%,70%, 80%, or 90% of a viral titer as compared to the viral titer priorto the five freeze/thaw cycles. In some cases, the pharmaceuticalcomposition, when administered to a patient with Leber's hereditaryoptic neuropathy, generates a higher average recovery of vision than acomparable pharmaceutical composition without the recombinant nucleicacid. In some cases, the pharmaceutical composition, when administeredto a patient with Leber's hereditary optic neuropathy, generates ahigher average recovery of vision than a comparable pharmaceuticalcomposition comprising a recombinant nucleic acid as set forth in SEQ IDNO: 15.

In another aspect, disclosed herein is a method of treating an eyedisorder, comprising administering any pharmaceutical compositiondisclosed herein to a patient in need thereof. In some cases, the eyedisorder is Leber's hereditary optic neuropathy (LHON). In some cases,the method comprises administering the pharmaceutical composition to oneor both eyes of the patient. In some cases, the pharmaceuticalcomposition is administered via intraocular or intravitreal injection.In some cases, the pharmaceutical composition is administered viaintravitreal injection. In some cases, about 0.01-0.1 mL of thepharmaceutical composition is administered via intravitreal injection.In some cases, about 0.05 mL of the pharmaceutical composition isadministered via intravitreal injection.

In some cases, the method further comprises administeringmethylprednisolone to the patient. In some cases, the methylprednisoloneis administered prior to the intravitreal injection of thepharmaceutical composition. In some cases, the methylprednisolone isadministered orally In some cases, the methylprednisolone isadministered daily for at least 1, 2, 3, 4, 5, 6, or 7 days prior to theintravitreal injection of the pharmaceutical composition. In some cases,the methylprednisolone is administered daily. In some cases, the a dailydosage of about 32 mg/60 kg methylprednisolone is administered. In somecases, the methylprednisolone is administered after the intravitrealinjection of the pharmaceutical composition. In some cases, the methodfurther comprises administering creatine phosphate sodium to thepatient. In some cases, the creatine phosphate sodium is administeredintravenously. In some cases, the methylprednisolone is administeredintravenously or orally. In some cases, the method comprisesadministering methylprednisolone intravenously for at least one day,which is followed by administering methylprednisolone orally for atleast a week. In some cases, the method comprises administeringmethylprednisolone intravenously for about 3 days, which is followed byadministering methylprednisolone orally for at least about 6 weeks. Insome cases, the methylprednisolone is administered intravenously at adaily dose of about 80 mg/60 kg. In some cases, the administering thepharmaceutical composition generates a higher average recovery of visionthan a comparable pharmaceutical composition without the recombinantnucleic acid. In some cases, the administering the pharmaceuticalcomposition generates a higher average recovery of vision than acomparable pharmaceutical composition comprising a recombinant nucleicacid as set forth in SEQ ID NO: 15.

INCORPORATION BY REFERENCE

All publications, patents, and patent applications mentioned in thisspecification are herein incorporated by reference to the same extent asif each individual publication, patent, or patent application wasspecifically and individually indicated to be incorporated by reference.

BRIEF DESCRIPTION OF THE DRAWINGS

The novel features of the invention are set forth with particularity inthe appended claims. A better understanding of the features andadvantages of the present invention will be obtained by reference to thefollowing detailed description that sets forth illustrative embodiments,in which the principles of the invention are utilized, and theaccompanying drawings of which:

FIG. 1 shows the PCR nucleic acid electrophoresis verification of ND4(lane A) and optimized ND4 (lane B) gene cloning results.

FIG. 2 shows the relative expression level comparison using qPCR betweenthe rAAV2-opt_ND4 (left black column) and rAAV2-ND4 (right blackcolumn). β-actin is the internal reference gene (white column).

FIG. 3 shows the relative expression level comparison usingimmunoblotting between the rAAV2-opt_ND4 (left black column) andrAAV2-ND4 (right black column). β-actin is the internal reference gene(white column).

FIG. 4 shows the fundus photographic results for rabbits injected withrAAV2-opt_ND4 (right) and rAAV2-ND4 (left), respectively.

FIG. 5 shows the fundus photographic results for a patient before (left)and after (right) the injection with rAAV2-optimized ND4.

FIG. 6 shows EGFP expression levels of rAAV2-ND4 (left) andrAAV2-opt_ND4* (right).

FIG. 7 shows the ND4 expression in 293T cells: rAAV2-ND4 (left) andrAAV2-opt_ND4* (right).

FIG. 8 shows the relative ND4 expression in 293T cells: rAAV2-ND4 (left)and rAAV2-opt_ND4* (right).

FIG. 9 shows the ND4 expression in rabbit optic nerve cells: rAAV2-ND4(left) and rAAV2-opt_ND4* (right).

FIG. 10 shows the relative ND4 expression in rabbit optic nerve cells:rAAV2-ND4 (left) and rAAV2-opt_ND4* (right).

FIG. 11 shows the fundus photographic results for rAAV2-ND4 (left) andrAAV2-opt_ND4* (right).

FIG. 12 shows the microscope inspection (HE staining) results forrAAV2-ND4 (left) and rAAV2-opt_ND4* (right).

FIG. 13 shows the fundus photographic results for rabbits injected withrAAV2-ND6 (A), rAAV-GFP (B) and PBS, respectively.

FIG. 14 shows the fundus photographic results for rabbits injected withrAAV2-opt_ND6 (A), rAAV2-ND6 (B), rAAV-EGFP (C), respectively.

FIG. 15 shows the relative ND6 expression in rabbit optic nerve cells:rAAV2-opt_ND6 (A), rAAV2-ND6 (B), and rAAV-EGFP (C).

FIG. 16 shows the relative ND6 expression by western blot: rAAV2-opt_ND6(A), rAAV2-ND6 (B), and rAAV-EGFP (C).

FIG. 17 shows the relative ND1 expression in rabbit optic nerve cells:rAAV2-opt_ND1 (A), rAAV2-ND1 (B), and rAAV-EGFP (C).

FIG. 18 shows the relative ND1 expression by western blot: rAAV2-opt_ND1(A), rAAV2-ND1 (B), and rAAV-EGFP (C).

DETAILED DESCRIPTION OF THE INVENTION Definitions

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of the ordinaryskill in the art to which this disclosure belongs. Although any methodsand materials similar or equivalent to those described herein can beused in the practice or testing of the formulations or unit dosesherein, some methods and materials are now described. Unless mentionedotherwise, the techniques employed or contemplated herein are standardmethodologies. The materials, methods and examples are illustrative onlyand not limiting.

As used herein and in the appended claims, the singular forms “a,”“and,” and “the” include plural referents unless the context clearlydictates otherwise. Thus, for example, reference to “a compound”includes a plurality of such agents, and reference to “the salt”includes reference to one or more salts (or to a plurality of salts) andequivalents thereof known to those skilled in the art, and so forth.

As used herein, unless otherwise indicated, the term “or” can beconjunctive or disjunctive. As used herein, unless otherwise indicated,any embodiment can be combined with any other embodiment.

As used herein, unless otherwise indicated, some inventive embodimentsherein contemplate numerical ranges. When ranges are present, the rangesinclude the range endpoints. Additionally, every subrange and valuewithin the range is present as if explicitly written out.

The term “about” and its grammatical equivalents in relation to areference numerical value and its grammatical equivalents as used hereincan include a range of values plus or minus 10% from that value, such asa range of values plus or minus 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or1% from that value. For example, the amount “about 10” includes amountsfrom 9 to 11.

The term “comprising” (and related terms such as “comprise” or“comprises” or “having” or “including”) is not intended to exclude thatin other certain embodiments, for example, an embodiment of anycomposition of matter, composition, method, or process, or the like,described herein, may “consist of” or “consist essentially of” thedescribed features.

The term “subject” refers to a mammal that has been or will be theobject of treatment, observation or experiment. The term “mammal” isintended to have its standard meaning, and encompasses humans, dogs,cats, sheep, and cows, for example. The methods described herein can beuseful in both human therapy and veterinary applications. In someembodiments, the subject is a human.

The term “treating” or “treatment” encompasses administration of atleast one compound disclosed herein, or a pharmaceutically acceptablesalt thereof, to a mammalian subject, particularly a human subject, inneed of such an administration and includes (i) arresting thedevelopment of clinical symptoms of the disease, such as cancer, (ii)bringing about a regression in the clinical symptoms of the disease,such as cancer, and/or (iii) prophylactic treatment for preventing theonset of the disease, such as cancer.

The term “therapeutically effective amount” of a chemical entitydescribed herein refers to an amount effective, when administered to ahuman or non-human subject, to provide a therapeutic benefit such asamelioration of symptoms, slowing of disease progression, or preventionof disease.

As used herein, unless otherwise indicated, the terms “nucleic acid” and“polynucleotide” can be used interchangeably.

Nucleic Acid and Polypeptide Sequences

Table 1 discloses all the nucleic acid and polypeptide sequencesdisclosed herein. The first column shows the SEQ ID NO of each sequence.The second column describes the nucleic acid or polypeptide construct.For example, the construct COX10-ND6-3′UTR is a nucleic acid combiningthe nucleic acid sequences of COX10 (SEQ ID NO: 1), ND6 (SEQ ID NO: 9),and 3′UTR (SEQ ID NO: 13) (from 5′ to 3′ without linker between thenucleic acid sequences.

TABLE 1 nucleic acid and polypeptide sequences and SEQ ID NOs SEQdescription sequence 1 COX10ATGGCCGCATCTCCGCACACTCTCTCCTCACGCCTCCTGACAGGTTGCGTAGGAGGCTCTGTCTGGTATCTTGAAAGAAGAACT 2 opt_COX10ATGGCCGCCTCTCCACACACACTGAGTAGCAGACTGCTGACCGGCTGTGTTGGCGGCTCTGTGTGGTATCTGGAACGGCGGACA 3 opt_COX10*ATGGCCGCCAGCCCCCACACCCTGAGCAGCCGCCTGCTGACCGGCTGCGTGGGCGGCAGCGTGTGGTACCTGGAGCGCCGCACC 4 COX8ATGTCCGTCCTGACGCGCCTGCTGCTGCGGGGCTTGACACGGCTCGGCTCGGCGGCTCCAGTGCGGCGCGCCAGAATCCATTCGTTG 5 CPA1GTGCTGCCCGCCTAGAAAGGGTGAAGTGGTTGTTTCCGTGACGGACTGAGTACGGGTGCCTGTCAGGCTCTTGCGGAAGTCCATGCGCCATTGGGAGGGCCTCGGCCGCGGCTCTGTGCCCTTGCTGCTGAGGGCCACTTCCTGGGTCATTCCTGGACCGGGAGCCGGGCTGGGGQTCACACGGGGGCTCCCGCGTGGCCGTCTCGGCGCCTGCGTGACCTCCCCGCCGGCGGGATGTGGCGACTACGTCGGGCCGCTGTGGC CTG 6ND4ATGCTAAAACTAATCGTCCCAACATTTATGTTACTACCACTGACATGGCTTTCCAAAAAACACATGATTTGGATCAACACAACCACCCACAGCCTAATTATTAGCATCATCCCTCTACTATTTTTTAACCAAATCAACAACAACCTATTTAGCTGTTCCCCAACCTTTTCCTCCGACCCCCTAACAACCCCCCTCCTAATGCTAACTACCTGGCTCCTACCCCTCACAATCATGGCAAGCCAAGGCCACTTATCCAGTGAACCACTATCACGAAAAAAACTCTACCTCTCTATGCTAATCTCCCTACAAATCTCCTTAATTATGACATTCACAGCCACAGAACTAATCATGTTTTATATCTTCTTCGAAACCACACTTATCCCCACCTTGGCTATCATCACCCGATGGGGCAACCAGCCAGAACGCCTGAACGCAGGCACATACTTCCTATTCTACACCCTAGTAGGCTCCCTTCCCCTACTCATCGCACTAATTTACACTCACAACACCCTAGGCTCACTAAACATTCTACTACTCACTCTGACTGCCCAAGAACTATCAAACTCCTGGGCCAACAACTTAATGTGGCTAGCTTACACAATGGCTTTTATGGTAAAGATGCCTCTTTACGGACTCCACTTATGGCTCCCTAAAGCCCATGTCGAAGCCCCCATCGCTGGGTCAATGGTACTTGCCGCAGTACTCTTAAAACTAGGCGGCTATGGTATGATGCGCCTCACACTCATTCTCAACCCCCTGACAAAACACATGGCCTACCCCTTCCTTGTACTATCCCTATGGGGCATGATTATGACAAGCTCCATCTGCCTACGAGAAACAGACCTAAAATCGCTCATTGCATACTCTTCAATGAGCCACATGGCCCTCGTAGTAACAGCCATTCTCATCCAAACCCCCTGGAGCTTCACCGGCGCAGTCATTCTCATGATCGCCCACGGGCTTACATCCTCATTACTATTCTGCCTAGCAAACTCAAACTACGAACGCACTCACAGTCGCATCATGATCCTCTCTCAAGGACTTCAAACTCTACTCCCACTAATGGCTTTTTGGTGGCTTCTAGCAAGCCTCGCTAACCTCGCCTTACCCCCCACTATTAACCTACTGGGAGAACTGTCTGTGQTAGTAACCACGTTCTCCTGGTGAAATATCACTCTCCTACTTACAGGACTCAACATGCTAGTCACAGCCCTATACTCCCTCTACATGTTTACCACAACACAATGGGGCTCACTCACCCACCACATTAACAACATGAAACCCTCATTCACACGAGAAAACACCCTCATGTTCATGCACCTATCCCCCATTCTCCTCCTATCCCTCAACCCCGACATCATTACCGGGTTTTCCTCTTAA 7 opt_ND4ATGCTGAAGCTGATCGTGCCCACCATCATGCTGCTGCCTCTGACCTGGCTGAGCAAGAAACACATGATCTGGATCAACACCACCACGCACAGCCTGATCATGAGCATCATCCGTGTGCTGTTCTTCAACCAGATCAACAACAACCTGTTCAGCTGCAGCCCCACCTTCAGCAGCGACCCTCTGACAACACCTCTGCTGATGCTGACCACCTGGCTGCTGCCCCTCACAATCATGGCCTCTCAGAGACACCTGAGCAGCGAGCCCCTGAGCCGGAAGAAACTGTACCTGAGCATGCTGATCTCCCTGCAGATCTCTCTGATCATGACCTTCACCGCCACCGAGCTGATCATGTTQTAGATCTTTTTCGAGAGAACGCTGATCCGCACACTGGCCATCATCACCAGATGGGGCAACCAGCCTGAGAGACTGAACGCCGGCACCTACTTTCTGTTCTACACCCTCGTGGGCAGCCTGCCACTGCTGATTGCCCTGATCTACACCCACAACACCCTGGGCTCCCTGAACATCCTGCTGCTGACACTGACAGCCCAAGAGCTGAGCAACAGCTGGGCCAACAATCTGATGTGGCTGGCCTACACAATGGCCTTCATGGTCAAGATGCCCCTGTACGGCCTGCACCTGTGGCTGCCTAAAGCTCATGTGGAAGCCCCTATCGCCGGCTCTATGGTGCTGGCTGCAGTGCTGCTGAAACTCGGCGGCTACGGCATGATGCGGCTGACCCTGATTCTGAATCCCCTGACCAAGCACATGGCCTATCCATTTCTGGTGCTGAGCCTGTGGGGCATGATTATGACCAGCAGCATCTGCCTGCGGCAGACCGATCTGAAGTCCCTGATCGCCTACAGCTCCATCAGCCACATGGCCCTGGTGGTCACCGCCATCCTGATTCAGACCCCTTGGAGCTTTACAGGCGCCGTGATCCTGATGATTGCCCACGGCCTGACAAGGAGCCTGGTGTTTTGTCTGGCCAACAGCAACTAGGAGCGGACCCACAGCAGAATCATGATCCTGTCTCAGGGCCTGCAGACCCTCCTGCCTCTTATGGCTTTTTGGTGGCTGCTGGCCTCTCTGGCCAATCTGGCACTGCCTCCTACCATCAATCTGCTGGGCGAGCTGAGCGTGCTGGTCACCACATTCAGCTGGTCCAATATCACCCTGCTGCTCACCGGCCTGAACATGCTGGTTACAGCCCTGTACTCCCTGTACATGTTCACCACCACACAGTGGGGAAGCCTGAGACACCACATCAACAATATGAAGCCCAGCTTCACCCGCGAGAACACCCTGATGTTCATGCATCTGAGCCCCATTCTGCTGCTGTCCCTGAATCCTGATATCATCACCGGCTTCTCCAGCTGA 8 opt_ND4*ATGCTGAAGCTGATCGTGCCCACCATCATGCTGCTGCCCCTGACCTGGCTGAGCAAGAAGCACATGATCTGGATCAACACCACCACCCACAGCCTGATCATCAGCATCATCCCCCTGCTGTTCTTCAACCAGATCAACAACAACCTGTTCAGCTGCAGCCCCACCTTCAGCAGCGACCCCCTGACCACCCCCCTGCTGATGCTGACCACCTGGCTGCTGCCCCTGACCATCATGGCCAGCCAGCGCCACCTGAGCAGCGAGCCCCTGAGCCGCAAGAAGCTGTACCTGAGCATGCTGATCAGCCTGCAGATCAGCCTGATCATGACCTTCACCGCCACCGAGCTGATCATGTTCTACATCTTCTTCGAGACCACCCTGATCCCCACCCTGGCCATCATCACCCGCTGGGGCAACCAGCCCGAGCGCCTGAACGCCGGCACCTACTTCCTGTTCTACACCCTGGTGGGCAGCCTGCCCCTGCTGATCGCCCTGATCTACACCCACAACACCCTGGGCAGCCTGAACATCCTGCTGCTGACCCTGACCGCCCAGGAGCTGAGCAACAGCTGGGCCAACAACCTGATGTGGCTGGCCTACACCATGGCCTTCATGGTGAAGATGCCCCTGTACGGCCTGCACCTGTGGCTGCCCAAGGCCCACGTGGAGGCCCCCATCGCCGGCAGCATGGTGCTGGCCGCCGTGCTGCTGAAGCTGGGCGGCTACGGCATGATGCGCCTGACCCTGATCCTGAACCCCCTGACCAAGCACATGGCCTACCCCTTCCTGGTGCTGAGCCTGTGGGGCATGATCATGACCAGCAGCATCTGCCTGCGCCAGACCGACCTGAAGAGCCTGATCGCCTACAGCAGCATCAGCCACATGGCCCTGGTGGTGACCGCCATCCTGATCCAGACCCCCTGGAGCTTCACCGGCGCCGTGATCCTGATGATCGCCCACGGCCTGACCAGCAGCCTGCTGTTCTGCCTGGCCAACAGCAACTACGAGCGCACCCACAGCCGCATCATGATCCTGAGCCAGGGCCTGCAGACCCTGCTGCCCCTGATGGCCTTCTGGTGGCTGCTTGGCCAGCCTGGCCAACCTGGCCCTGCCCCCCACCATCAACCTGCTGGGCGAGCTGAGCGTGCTGGTGACCACCTTCAGCTGGAGCAACATCACCCTGCTGCTGACCGGCCTGAACATGCTGGTGACCGCCCTGTACAGCCTGTACATGTTCACCACCACCCAGTGGGGCAGCCTGACCCACCACATCAACAACATGAAGCCCAGCTTCACCCGCGAGAACACCCTGATGTTCATGCACCTGAGCCCCATCCTGCTGCTGAGCCTGAACCCCGACATCATCACCGGCTTCAGCAGCTAA 9 ND6ATGATGTATGCTTTGTTTCTGTTGAGTGTGGGTTTAGTAATGGGGTTTGTGGGGTTTTCTTCTAAGCCTTCTCCTATTTATGGGGGTTTAGTATTGATTGTTAGCGGTGTGGTCGGGTGTGTTATTATTCTGAATTTTGGGGGAGGTTATATGGGTTTAATGGTTTTTTTAATTTATTTAGGGGGAATGATGGTTGTCTTTGGATATACTACAGCGATGGCTATTGAGGAGTATCCTGAGGCATGGGGGTCAGGGGTTGAGGTCTTGGTGAGTGTTTTAGTGGGGTTAGCGATGGAGGTAGGATTGGTGCTGTGGGTGAAAGAGTATGATGGGGTGGTGGTTGTGGTAAACTTTAATAGTGTAGGAAGCTGGATGATTTATGAAGGAGAGGGGTCAGGGTTGATTCGGGAGGATCCTATTGGTGCGGGGGCTTTGTATGATTATGGGCGTTTGGTTAGTAGTAGTTACTGGTTGGACATTGTTTGTTGGTGTATATATTGTAATTGAGATTGCTCGGGGGAATTAG 10 opt_ND6ATGATGTACGCCCTGTTCCTGCTGAGCGTGGGCCTGGTGATGGGCTTCGTGGGCTTCAGCAGCAAGCCCAGCCCCATCTACGGCGGCCTGGTGCTGATCGTGAGCGGCGTGGTGGGCTGCGTGATCATCCTGAACTTCGGCGGCGGCTACATGGGCCTGATGGTGTTCCTGATCTACCTGGGCGGCATGATGGTGGTGTTCGGCTACACCACCGCCATGGCCATCGAGGAGTACCCCGAGGCCTGGGGCAGCGGCGTGGAGGTGCTGGTGAGCGTGCTGGTGGGCCTGGCCATGGAGGTGGGCCTGGTGCTGTGGGTGAAGGAGTACGACGGCGTGGTGGTGGTGGTGAACTTCAACAGCGTGGGCAGCTGGATGATCTACGAGGGCGAGGGCAGCGGCCTGATCCGCGAGGACCCCATCGGCGCCGGCGCCCTGTACGACTACGGCCGCTGGCTGGTGGTGGTGACCGGCTGGACCCTGTTCGTGGGCGTGTACATCGTGATCGAGATCGCCCGCGGCAACTAA 11 ND1ATGGCCAACCTCCTACTCCTCATTGTACCCATTCTAATCGCAATGGCATTCCTAATGCTTACCGAACGAAAAATTCTAGGCTATATGCAACTACGCAAAGGCCCCAACGTTGTAGGCCCCTACGGGCTACTACAACCCTTCGCTGACGCCATAAAACTCTTCACCAAAGAGCCCCTAAAACCCGCCACATCTACCATCACCCTCTACATCACCGCCCCGACCTTAGCTCTCACCATCGCTCTTCTACTATGGACCCCCCTCCCCATGCCCAACCCCCTGGTCAACCTCAACCTAGGCCTCCTATTTATTCTAGCCACCTCTAGCCTAGCCGTTTACTCAATCCTCTGGTCAGGGTGGGCATCAAACTCAAACTACGCCCTGATCGGCGCACTGCGAGCAGTAGCCCAAACAATCTCATATGAAGTCACCCTAGCCATCATTCTACTATCAACATTACTAATGAGTGGCTCCTTTAACCTCTCCACCCTTATCACTAACACAAGAACACCTCTGGTTACTCCTGCCATDATGGCCCTTGGCCATGATGTGGTTTATCTCCACACTAGCAGAGACCAACCGAACCCCCTTCGACCTTGCCGAAGGGGAGTCCGAACTAGTCTCAGGCTTCAACATCGAATACGCCGCAGGCCCCTTCGCCCTATTCTTCATGGCCGAATACACAAACATTATTATGATGAACACCCTCACCACTACAATCTTCCTAGGAACAACATATGACGCACTCTCCCCTGAACTCTACACAACATATTTTGTCACCAAGACCCTACTTCTAACCTCCCTGTTCTTATGGATTCGAACAGCATACCCCCGATTCCGCTACGACCAACTCATGCACCTCCTATGGAAAAACTTCCTACCACTCACCCTAGCATTACTTATGTGGTATGTCTCCATGCCCATTACAATCTCCAGCATTCCCCCTCAAACCTAA 12opt_ND1ATGGCCAACCTGCTGCTGCTGATCGTGCCCATCCTGATCGCCATGGCCTTCCTGATGCTGACCGAGCGCAAGATCCTGGGCTACATGCAGCTGCGCAAGGGCCCCAACGTGGTGGGCCCCTACGGCCTGCTGCAGCCCTTCGCCGACGCCATCAAGCTGTTCACCAAGGAGCCCCTGAAGCCCGCCACCAGCACCATCACCCTGTACATCACCGCCCCCACCCTGGCCCTGACCATCGCCCTGCTGCTGTGGACCCCCCTGCCCATGCCCAACCCCCTGGTGAACCTGAACCTGGGCCTGCTGTTCATCCTGGCCACCAGCAGCCTGGCCGTGTACAGCATCCTGTGGAGCGGCTGGGCCAGCAACAGCAACTACGCCCTGATCGGCGCCCTGCGCGCCGTGGCCCAGACCATCAGCTACGAGGTGACCCTGGCCATCATCCTGCTGAGCACCCTGCTGATGAGCGGCAGCTTCAACCTGAGCACCCTGATCACCACCCAGGAGCACCTGTGGCTGCTGCTGCCCAGCTGGCCCCTGGCCATGATGTGGTTCATCAGCACCCTGGCCGAGACCAACCGCACCCCCTTCGACCTGGCCGAGGGCGAGAGCGAGCTGGTGAGCGGCTTCAACATCGAGTACGCCGCCGGCCCCTTCGCCCTGTTCTTCATGGCCGAGTACACCAACATCATCATGATGAACACCCTGACCACCACCATCTTCCTGGGCACCACCTACGACGCCCTGAGCCCCGAGCTGTACACCACCTACTTCGTGACCAAGACCCTGCTGCTGACCAGCCTGTTCCTGTGGATCCGCACCGCCTACCCCCGCTTCCGCTACGACCAGCTGATGCACCTGCTGTGGAAGAACTTCCTGCCCCTGACCCTGGCCCTGCTGATGTGGTACGTGAGCATGCCCATCACCATCAGCAGCATCCCCCCCCAGACCTAA 13 3′UTRGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCAGCCCCTGTCCTCCCTTCACCCCCATTGCGTATGAGCATTTCAGAACTCCAAGGAGTCACAGGCATCTTTATAGTTCACGTTAACATATAGACACTGTTGGAAGCAGTTCCTTCTAAAAGGGTAGCCCTGGACTTAATACCAGCCGGATACCTCTGGCCCCCACCCCATTACTGTACCTCTGGAGTCACTACTGTGGGTCGCCACTCCTCTGCTACACAGCACGGCTTTTTCAAGGCTGTATTGAGAAGGGAAGTTAGGAAGAAGGGTGTGCTGGGCTAACCAGCCCACAGAGCTCACATTCCTGTCCCTTGGGTGAAAAATACATGTCCATCCTGATATCTCCTGAATTCAGAAATTAGCCTCCACATGTGCAATGGCTTTAAGAGCCAGAAGCAGGGTTCTGGGAATTTTGCAAGTTACCTGTGGCCAGGTGTGGTCTCGGTTACCAAATACGGTTACCTGCAGCTTTTTAGTCCTTTGTGCTCCCACGGGTCTACAGAGTCCCATCTGCCCAAAGGTCTTGAAGCTTGACAGGATGTTTTCGATTACTCAGTCTCCCAGGGCACTACTGGTCCGTAGGATTCGATTGGTCGGGGTAGGAGAGTTAAACAACATTTAAACAGAGTTCTCTCAAAAATGTCTAAAGGGATTGTAGGTAGATAACATCCAATCACTGTTTGCACTTATCTGAAATCTTCCCTCTTGGCTGCCCCCAGGTATTTACTGTGGAGAACATTGCATAGGAATGTCTGGAAAAAGCTTCTACAACTTGTTACAGCCTTCACATTTGTAGAAGCTTT 14 3′UTR*GAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCA 15 COX10-ND4-3′UTRATGGCCGCATCTCCGCACACTCTCTCCTCACGCCTCCTGACAGGTTGCGTAGGAGGCTCTGTCTGGTATCTTGAAAGAAGAACTATGCTAAAACTAATCGTCCCAACAATTATGTTACTACCACTGACATGGCTTTCCAAAAAACACATGATTTGGATCAACACAACCACCCACAGCCTAATTATTAGCATCATCCCTCTACTATTTTTTAACCAAATCAACAACAACCTATTTAGCTGTTCCCCAACCTTTTCCTCCGACCCCCTAACAACCCCCCTCCTAATGCTAACTACCTGGCTCCTACCCCTCACAATCATGGCAAGCCAACGCCACTTATCCAGTGAACCACTATCACGAAAAAAACTCTACCTCTCTATGCTAATCTCCCTACAAATCTCCTTAATTATGACATTCACAGCCACAGAACTAATCATGTTTTATATCTTCTTCGAAACCACACTTATCCCCACCTTGGCTATCATCACCCGATGGGGCAAACCAGCCAGAACGCCTGAACGCAGGCACATACTTCCTATTCTACACCCTAGTAGGCTCCCTTCCCCTACTCATCGCACTAATTTACACTCACAACACCCTAGGCTCACTAAACATTCTACTACTCACTCTCACTGCCCAAGAACTATCAAACTCCTGGGCCAACAACTTAATGTGGCTAGCTTACACAATGGCTTTTATGGTAAAGATGCCTCTTTACGGACTCCACTTATGGCTCCCTAAAGCCCATGTCGAAGCCCCCATCGCTGGGTCAATGGTACTTGCCGCAGTACTCTTAAAACTAGGCGGCTATGGTATGATGCGCCTCACACTCATTCTCAACCCCCTGACAAAACACATGGCCTACCCCTTCCTTGTACTATCCCTATGGGGCATGATTATGACAAGCTCCATCTGCCTACGACAAACAGACCTAAAATCGCTCATTGCATACTCTTCAATCAGCCACATGGCCCTCGTAGTAACAGCCATTCTCATCCAAACCCCCTGGAGCTTCACCGGCGCAGTCATTCTCATGATCGCCCACGGGCTTACATCCTCATTACTATTCTGCCTAGCAAACTCAAACTACGAACGCACTCACAGTCGCATCATGATCCTCTCTCAAGGACTTCAAACTCTACTCCCACTAATGGCTTTTTGGTGGCTTCTAGCAAGCCTCGCTAACCTCGCCTTACCCCCCACTATTAACCTACTGGGAGAACTCTCTGTGCTAGTAACCACGTTCTCCTGGTCAAATATCACTCTCCTACTTACAGGACTCAACATGCTAGTCACAGCCCTATACTCCCTCTACATGTTTACCACAACACAATGGGGCTCACTCACCCACCACATTAACAACATGAAACCCTCATTCACACGAGAAAACACCCTCATGTTCATGCACCTATCCCCCATTCTCCTCCTATCCCTCAACCCCGACATCATTACCGGGTTTTCCTCTTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCAGCCCCTGTCCTCCCTTCACCCCCATTGCGTATGAGCATTTCAGAACTCCAAGGAGTCACAGGCATCTTTATAGTTCACGTTAACATATAGACACTGTTGGAAGCAGTTCCTTCTAAAAGGGTAGCCCTGGACTTAATACCAGCCGGATACCTCTGGCCCCCACCCCATTACTGTACCTCTGGAGTCACTACTGTGGGTCGCCACTCCTCTGCTACACAGCACGGCTTTTTCAAGGCTGTATTGAGAAGGGAAGTTAGGAAGAAGGGTGTGCTGGGCTAACCAGCCCACAGAGCTCACATTCCTGTCCCTTGGGTGAAAAATACATGTCCATCCTGATATCTCCTGAATTCAGAAATTAGCCTCCACATGTGCAATGGCTTTAAGAGCCAGAAGCAGGGTTCTGGGAATTTTGCAAGTTACCTGTGGCCAGGTGTGGTCTCGGTTACCAAATACGGTTACCTGCAGCTTTTTAGTCCTTTGTGCTCCCACGGGTCTACAGAGTCCCATCTGCCCAAAGGTCTTGAAGCTTGACAGGATGTTTTCGATTACTCAGTCTCCCAGGGCACTACTGGTCCGTAGGATTCGATTGGTCGGGGTAGGAGAGTTAAACAACATTTAAACAGAGTTCTCTCAAAAATGTCTAAAGGGATTGTAGGTAGATAACATCCAATCACTGTTTGCACTTATCTGAAATCTTCCCTCTTGGCTGCCCCCAGGTATTTACTGTGGAGAACATTGCATAGGAATGTCTGGAAAAAGCTTCTACAACTTGTTACAGCCTTCACATTTGTAGAAGCTTT 16 COX10-ND4-3′UTR*ATGGCCGCATCTCCGCACACTCTCTCCTCACGCCTCCTGACAGGTTGCGTAGGAGGCTCTGTCTGGTATCTTGAAAGAAGAACTATGCTAAAACTAATCGTCCCCAATTATGTTACTACCACTGACATGGCTTTCCAAAAAACACATGATTTGGATCAACACAACCACCCACAGCCTAATTATTAGCATCATCCCTCTACTATTTTTTAACCAAATCAACAACAACCTATTTAGCTGTTCCCCAACCTTTTCCTCCGACCCCCTAACAACCCCCCTCCTAATGCTAACTACCTGGCTCCTACCCCTCACAATCATGGCAAGCCAACGCCACTTATCCAGTGAACCACTATCACGAAAAAAACTCTACCTCTCTATGCTAATCTCCCTACAAATCTCCTTAATTATGACATTCACAGCCACAGAACTAATCATGTTTTATATCTTCTTCGAAACCACACTTATCCCCACCTTGGCTATCATCACCCGATGGGGCAACCAGCCAGAACGCCTGAACGCAGGCACATACTTCCTATTCTACACCCTAGTAGGCTCCCTTCCCCTACTCATCGCACTAATTTACACTCACAACACCCTAGGCTCACTAAACATTCTACTACTCACTCTCACTGCCCAAGAACTATCAAACTCCTGGGCCAACAACTTAATGTGGCTAGCTTACACAATGGCTTTTATGGTAAAGATGCCTCTTTACGGACTCCACTTATGGCTCCCTAAAGCCCATGTCGAAGCCCCCATCGCTGGGTCAATGGTACTTGCCGCAGTACTCTTAAAACTAGGCGGCTATGGTATGATGCGCCTCACACTCATTCTCAACCCCCTGACAAAACACATGGCCTACCCCTTCCTTGTACTATCCCTATGGGGCATGATTATGACAAGCTCCATCTGCCTACGACAAAACAGACCTAAAUCGCTCATTGCATACTCTTCAATCAGCCACATGGCCCTCGTAGTAACAGCCATTCTCATCCAAACCCCCTGGAGCTTCACCGGCGCAGTCATTCTCATGATCGCCCACGGGCTTACATCCTCATTACTATTCTGCCTAGCAAACTCAAACTACGAACGCACTCACAGTCGCATCATGATCCTCTCTCAAGGACTTCAAACTCTACTCCCACTAATGGCTTTTTGGTGGCTTCTAGCAAGCCTCGCTAACCTCGCCTTACCCCCCACTATTAACCTACTGGGAGAACTCTCTGTGCTAGTAACCACGTTCTCCTGGTCAAATATCACTCTCCTACTTACAGGACTCAACATGCTAGTCACAGCCCTATACTCCCTCTACATGTTTACCACAACACAATGGGGCTCACTCACCCACCACATTAACAACATGAAACCCTCATTCACACGAGAAAACACCCTCATGTTCATGCACCTATCCCCCATTCTCCTCCTATCCCTCAACCCCGACATCATTACCGGGTTTTCCTCTTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCA 17 COX10-opt_ND4-3′UTRATGGCCGCATCTCCGCACACTCTCTCCTCACGCCTCCTGACAGGTTGCGTAGGAGGCTCTGTCTGGTATCTTGAAAGAAGCTATGCTGAAGCTGATCGTGCCCACCATCATGCTGCTGCCTCTGACCTGGCTGAGCAAGAAACACATGATCTGGATCAACACCACCACGCACAGCCTGATCATCAGCATCATCCCTCTGCTGTTCTTCAACCAGATCAACAACAACCTGTTCAGCTGCAGCCCCACCTTCAGCAGCGACCCTCTGACAACACCTCTGCTGATGCTGACCACCTGGCTGCTGCCCCTCACAATCATGGCCTCTCAGAGACACCTGAGCAGCGAGCCCCTGAGCCGGAAGAAACTGTACCTGAGCATGCTGATCTCCCTGCAGATCTCTCTGATCATGACCTTCACCGCCACCGAGCTGATCATGTTCTACATCTTTTTCGAGACAACGCTGATCCCCACACTGGCCATCATCACCAGATGGGGCAACCAGCCTGAGAGACTGAACGCCGGCACCTACTTTCTGTTCTACACCCTCGTGGGCAGCCTGCCACTGCTGATTGCCCTGATCTACACCCACAACACCCTGGGCTCCCTGAACATCCTGCTGCTGACACTGACAGCCCAAGAGCTGAGCAACAGCTGGGCCAACAATCTGATGTGGCTGGCCTACACAATGGCCTTCATGGTCAAGATGCCCCTGTACGGCCTGCACCTGTGGCTGCCTAAAGCTCATGTGGAAGCCCCTATCGCCGGCTCTATGGTGCTGGCTGCAGTGCTGCTGAAACTCGGCGGCTACGGCATGATGCGGCTGACCCTGATTCTGAATCCCCTGACCAAGCACATGGCCTATCCATTTCTGGTGCTGAGCCTGTGGGGCATGATTATGACCAGCAGCATCTGCCTGCGGCAGACCGATCTGAAGTCCCTGATCGCCTACAGCTCCATCAGCCACATGGCCCTGGTGGTCACCGCCATCCTGATTCAGACCCCTTGGAGCTTTACAGGCGCCGTGATCCTGATGATTGCCCACGGCCTGACAAGCAGCCTGCTGTTTTGTCTGGCCAACAGCAACTACGAGCGGACCCACAGCAGAATCATGATCCTGTCTCAGGGCCTGCAGACCCTCCTGCCTCTTATGGCTTTTTGGTGGCTGCTGGCCTCTCTGGCCAATCTGGCACTGCCTCCTACCATCAAlCTGGTGGGCGAGCTGAGCGTGCTGGTCACCACATTCAGCTGGTCCAATATCACCCTGCTGCTCACCGGCCTGAACATGCTGGTTACAGCCCTGTACTCCCTGTACATGTTCACCACCACACAGTGGGGAAGCCTGACACACCACATCAACAATATGAAGCCCAGCTTCACCCGCGAGAACACCCTGATGTTCATGCATCTGAGCCCCATTCTGCTGCTGFCCCTGAATCCTGATATCATCACCGGCTTCTCCAGCTGAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGGTGGACTGCCAGCCCCTGTCCTCCCTTCACCCCCATTGCGTATGAGCATTTCAGCTCCAAGGAGTCACAGGCATCTTTATAGTTCACGTTAACATATAGACACTGTTGGAAGCAGTTCCTTCTAAAAGGGTAGCCCTGGACTTAATACCAGCCGGATACCTCTGGCCCCCACCCCATTACTGTACCTCTGGAGTCACTACTGTGGGTCGCCACTCCTCTGCTACACAGCACGGCTTTTTCAAGGCTGTATTGAGAAGGGAAGTTAGGAAGAAGGGTGTGGTGGGCTAACCAGCCCACAGAGCTCACATTCCTGTCCCTTGGGTGAAAAATACATGTCCATCCTGATATCTCCTGAATTCAGAAATTAGCCTCCACATGTGCAATGGCTTTAAGAGCCAGAAGCAGGGTTCTGGGAATTTTGCAAGTTACCTGTGGCCAGGTGTGGTCTCGGTTACCAAATACGGTTACCTGCAGCTTTTTAGTCCTTTGTGCTCCCACGGGTCTACAGAGTCCCATCTGCCCAAAGGTCTTGAAGCTTGACAGGATGTTTTCGATTACTCAGTCTCCCAGGGCACTACTGGTCCGTAGGATTCGATTGGTCGGGGTAGGAGAGTTAAACAACATTTAAACAGAGTTCTCTCAAAAATGTCTAAAGGGATTGTAGGTAGATAACATCCAATCACTGTTTGCACTTATCTGAAATCTTCCCTCTTGGCTGCCCCCAGGTATTTACTGTGGAGAACATTGCATAGGAATGTCTGGAAAAAGCTTCTACAACTTGTTACAGCCTTCACATTTGTAGAAGCTTT 18 COX10-opt_ND4-3′UTRATGGCCGCATCTCCGCACACTCTCTCCTCACGCCTCCTGACAGGTTGCGTAGGAGGCTCTGTCTGGTATCTTGAAAGAAAACTATGCTGAAGCTGATCGTGCCCACCATCATGCTGCTGCCTCTGACCTGGCTGAGCAAGAAACACATGATCTGGATCAACACCACCACGCACAGCCTGATCATCAGCATCATCCCTCTGCTGTTCTTCAACCAGATCAACAACAACCTGTTCAGCTGCAGCCCCACCTTCAGCAGCGACCCTCTGACAACACCTCTGCTGATGCTGACCACCTGGCTGCTGCCCCTCACAATCATGGCCTCTCAGAGACACCTGAGCAGCGAGCCCCTGAGCCGGAAGAAACTGTACCTGAGCATGCTGATCTCCCTGCAGATCTCTCTGATCATGACCTTCACCGCCACCGAGCTGATCATGTTCTACATCTTTTTCGAGACAACGCTGATCCCCACACTGGCCATCATCACCAGATGGGGCAACCAGCCTGAGAGACTGAACGCCGGCACCTACTTTCTGTTCTACACCCTCGTGGGCAGCCTGCCACTGCTGATTGCCCTGATCTACACCCACAACACCCTGGGCTCCCTGAACATCCTGCTGCTGACACTGACAGCCCAAGAGCTGAGCAACAGCTGGGCCAACAATCTGATGTGGCTGGCCTACACAATGGCCTTCATGGTCAAGATGCCCCTGTACGGCCTGCACCTGTGGCTGCCTAAAGCTCATGTGGAAGCCCCTATCGCCGGCTCTATGGTGCTGGCTGCAGTGCTGCTGAAACTCGGCGGCTACGGCATGATGCGGCTGACCCTGATTCTGAATCCCCTGACCAAGCACATGGCCTATCCATTTCTGGTGCTGAGCCTGTGGGGCATGATTATGACCAGCAGCATCTGCCTGCGGCAGACCGATCTGAAGTCCCTGATCGCCTACAGCTCCATCAGCCACATGGCCCTGGTGGTCACCGCCATCCTGATTCAGACCCCTTGGAGCTTTACAGGCGCCGTGATCCTGATGATTGCCCACGGCCTGACAAGCAGCCTGCTGTTTTGTCTGGCCAACAGCAACTACGAGCGGACCCACAGCAGAATCATGATCCTGTCTCAGGGCCTGCAGACCCTCCTGCCTCTTATGGCTTTTTGGTGGCTGCTGGCCTCTCTGGCCAATCTGGCACTGCCTCCTACCATCAATCTGCTGGGCGAGCTGAGCGTGCTGGTCACCACATTCAGCTGGTCCAATATCACCCTGCTGCTCACCGGCCTGAACATGCTGGTTACAGCCCTGTACTCCCTGTACATGTTCACCACCACACAGTGGGGAAGCCTGACACACCACATCAACAATATGAAGCCCAGCTTCACCCGCGAGAACACCCTGATGTTCATGCATCTGAGCCCCATTCTGCTGCTGTCCCTGAATCCTGATATCATCACCGGCTTCTCCAGCTGAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCA 19COX10-opt_ND4*-3′UTRATGGCCGCATCTCCGCACACTCTCTCCTCACGCCTCCTGACAGGTTGCGTAGGAGGCTCTGTCTGGTATCTTGAAAGAAAACTATGCTGAAGCTGATCGTGCCCACCATCATGCTGCTGCCCCTGACCTGGCTGAGCAAGAAGCACATGATCTGGATCAACACCACCACCCACAGCCTGATCATCAGCATCATCCCCCTGCTGTTCTTCAACCAGATCAACAACAACCTGTTCAGCTGCAGCCCCACCTTCAGCAGCGACCCCCTGACCACCCCCCTGCTGATGCTGACCACCTGGCTGCTGCCCCTGACCATCATGGCCAGCCAGCGCCACCTGAGCAGCGAGCCCCTGAGCCGCAAGAAGCTGTACCTGAGCATGCTGATCAGCCTGCAGATCAGCCTGATCATGACCTTCACCGCCACCGAGCTGATCATGTTCTACATCTTCTTCGAGACCACCCTGATCCCCACCCTGGCCATCATCACCCGCTGGGGCAACCAGCCCGAGCGCCTGAACGCCGGCACCTACTTCCTGTTCTACACCCTGGTGGGCAGCCTGCCCCTGCTGATCGCCCTGATCTACACCCACAACACCCTGGGCAGCCTGAACATCCTGCTGCTGACCCTGACCGCCCAGGAGCTGAGCAACAGCTGGGCCAACAACCTGATGTGGCTGGCCTACACCATGGCCTTCATGGTGAAGATGCCCCTGTACGGCCTGCACCTGTGGCTGCCCAAGGCCCACGTGGAGGCCCCCATCGCCGGCAGCATGGTGCTGGCCGCCGTGCTGCTGAAGCTGGGCGGCTACGGCATGATGCGCCTGACCCTGATCCTGAACCCCCTGACCAAGCACATGGCCTACCCCTTCCTGGTGCTGAGCCTGTGGGGCATGATCATGACCAGCAGCATCTGCCTGCGCCAGACCGACCTGAAGAGCCTGATCGCCTACAGCAGCATCAGCCACATGGCCCTGGTGGTGACCGCCATCCTGATCCAGACCCCCTGGAGCTTCACCGGCGCCGTGATCCTGATGATCGCCCACGGCCTGACCAGCAGCCTGCTGTTCTGCCTGGCCAACAGCAACTACGAGCGCACCCACAGCCGCATCATGATCCTGAGCCAGGGCCTGCAGACCCTGCTGCCCCTGATGGCCTTCTGGTGGCTGCTGGCCAGCCTGGCCAACCTGGCCCTGCCCCCCACCATCAACCTGCTGGGCGAGCTGAGCGTGCTGGTGACCACCTTCAGCTGGAGCAACATCACCCTGCTGCTGACCGGCCTGAACATGCTGGTGACCGCCCTGTACAGCCTGTACATGTTCACCACCACCCAGTGGGGCAGCCTGACCCACCACATCAACAACATGAAGCCCAGCTTCACCCGCGAGAACACCCTGATGTTCATGCACCTGAGCCCCATCCTGCTGCTGAGCCTGAACCCCGACATCATCACCGGCTTCAGCAGCTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGGTGGACTGCCAGCCCCTGTCCTCCCTTCACCCCCATTGCGTATGAGCATTTCAGAACTCCAAGGAGTCACAGGCATCTTTATAGTTCACGTTAACATATAGACACTGTTGGAAGCAGTTCCTTCTAAAAGGGTAGCCCTGGACTTAATACCAGCCGGATACCTCTGGCCCCCACCCCATTACTGTACCTCTGGAGTCACTACTGTGGGTCGCCACTCCTCTGCTACACAGCACGGCTTTTTCAAGGCTGTATTGAGAAGGGAAGTTAGGAAGAAGGGTGTGCTGGGCTAACCAGCCCACAGAGCTCACATTCCTGTCCCTTGGGTGAAAAATACATGTCCATCCTGATATCTCCTGAATTCAGAAATTAGCCTCCACATGTGCAATGGCTTTAAGAGCCAGAAGCAGGGTTCTGGGAATTTTGCAAGTTACCTGTGGCCAGGTGTGGTCTCGGTTACCAAATACGGTTACCTGCAGCTTTTTAGTCCTTTGTGCTCCCACGGGTCTACAGAGTCCCATCTGCCCAAAGGTCTTGAAGCTTGACAGGATGTTTTCGATTACTCAGTCTCCCAGGGCACTACTGGTCCGTAGGATTCGATTGGTCGGGGTAGGAGAGTTAAACAACATTTAAACAGAGTTCTCTCAAAAATGTCTAAAGGGATTGTAGGTAGATAACATCCAATCACTGTTTGCACTTATCTGAAATCTTCCCTCTTGGCTGCCCCCAGGTATTTACTGTGGAGAACATTGCATAGGAATGTCTGGAAAAAGCTTCTACAACTTGTTACAGCCTTCACATTTGTAGAAGCTTT 20 COX10-opt_ND4*-3′UTR*ATGGCCGCATCTCCGCACACTCTCTCCTCACGCCTCCTGACAGGTTGCGTAGGAGGCTCTGTCTGGTATCTTGAAAGAAGAACTATGCTGAAGCTGATCGTGCCCACCATCATGCTGCTGCCCCTGACCTGGCTGAGCAAGAAGCACATGATCTGGATCAACACCACCACCCACAGCCTGATCATCAGCATCATCCCCCTGCTGTTCTTCAACCAGATCAACAACAACCTGTTCAGCTGCAGCCCCACCTTCAGCAGCGACCCCCTGACCACCCCCCTGCTGATGCTGACCACCTGGCTGCTGCCCCTGACCATCATGGCCAGCCAGCGCCACCTGAGCAGCGAGCCCCTGAGCCGCAAGAAGCTGTACCTGAGCATGCTGATCAGCCTGCAGATCAGCCTGATCATGACCTTCACCGCCACCGAGCTGATCATGTTCTACATCTTCTTCGAGACCACCCTGATCCCCACCCTGGCCATCATCACCCGCTGGGGCAACCAGCCCGAGCGCCTGAACGCCGGCACCTACTTCCTGTTCTACACCCTGGTGGGCAGCCTGCCCCTGCTGATCGCCCTGATCTACACCCACAACACCCTGGGCAGCCTGAACATCCTGCTGCTGACCCTGACCGCCCAGGAGCTGAGCAACAGCTGGGCCAACAACCTGATGTGGCTGGCCTACACCATGGCCTTCATGGTGAAGATGCCCCTGTACGGCCTGCACCTGTGGCTGCCCAAGGCCCACGTGGAGGCCCCCATCGCCGGCAGCATGGTGCTGGCCGCCGTGCTGCTGAAGCTGGGCGGCTACGGCATGATGCGCCTGACCCTGATCCTGAACCCCCTGACCAAGCACATGGCCTACCCCTTCCTGGTGCTGAGCCTGTGGGGCATGATCATGACCAGCAGCATCTGCCTGCGCCAGACCGACCTGAAGAGCCTGATCGCCTACAGCAGCATCAGCCACATGGCCCTGGTGGTGACCGCCATCCTGATCCAGACCCCCTGGAGCTTCACCGGCGCCGTGATCCTGATGATCGCCCACGGCCTGACCAGCAGCCTGCTGTTCTGCCTGGCCAACAGCAACTACGAGCGCACCCACAGCCGCATCATGATCCTGAGCCAGGGCCTGCAGACCCTGCTGCCCCTGATGGCCTTCTGGTGGCTGCTGGCCAGCCTGGCCAACCTGGCCCTGCCCCCCACCATCAACCTGCTGGGCGAGCTGAGCGTGCTGGTGACCACCTTCAGCTGGAGCAACATCACCCTGCTGCTGACCGGCCTGAACATGCTGGTGACCGCCCTGTACAGCCTGTACATGTTCACCACCACCCAGTGGGGCAGCCTGACCCACCACATCAACAACATGAAGCCCAGCTTCACCCGCGAGAACACCCTGATGTTCATGCACCTGAGCCCCATCCTGCTGCTGAGCCTGAACCCCGACATCATCACCGGCTTCAGCAGCTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCA 21COX10-ND6-3′UTRATGGCCGCATCTCCGCACACTCTCTCCTCACGCCTCCTGACAGGTTGCGTAGGAGGCTCTGTCTGGTATCTTGAAAGAAGAACTATGATGTATGCTTTGTTTCTGTTGAGTGTGGGTTTAGTAATGGGGTTTGTGGGGTTTTCTTCTAAGCCTTCTCCTATTTATGGGGGTTTAGTATTGATTGTTAGCGGTGTGGTCGGGTGTGTTATTATTCTGAATTTTGGGGGAGGTTATATGGGTTTAATGGTTTTTTTAATTTATTTAGGGGGAATGATGGTTGTCTTTGGATATACTACAGCGATGGCTATTGAGGAGTATCCTGAGGCATGGGGGTCAGGGGTTGAGGTCTTGGTGAGTGTTTTAGTGGGGTTAGCGATGGAGGTAGGATTGGTGCTGTGGGTGAAAGAGTATGATGGGGTGGTGGTTGTGGTAAACTTTAATAGTGTAGGAAGCTGGATGATTTATGAAGGAGAGGGGTCAGGGTTGATTCGGGAGGATCCTATTGGTGCGGGGGCTTTGTATGATTATGGGCGTTGGTTAGTAGTAGTTACTGGTTGGACATTGTTTGTTGGTGTATATATTGTAATTGAGATTGCTCGGGGGAATTAGGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCAGCCCCTGTCCTCCCTTCACCCCCATTGCGTATGAGCATTTCAGAACTCCAAGGAGTCACAGGCATCTTTATAGTTCACGTTAACATATAGACACTGTTGGAAGCAGTTCCTTCTAAAAGGGTAGCCCTGGACTTAATACCAGCCGGATACCTCTGGCCCCCACCCCATTACTGTACCTCTGGAGTCACTACTGTGGGTCGCCACTCCTCTGCTACACAGCACGGCTTTTTCAAGGCTGTATTGAGAAGGGAAGTTAGGAAGAAGGGTGTGCTGGGCTAACCAGCCCACAGAGCTCACATTCCTGTCCCTTGGGTGAAAAATACATGTCCATCCTGATATCTCCTGAATTCAGAAATTAGCCTCCACATGTGCAATGGCTTTAAGAGCCAGAAGCAGGGTTCTGGGAATTTTGCAAGTTACCTGTGGCCAGGTGTGGTCTCGGTTACCAAATACGGTTACCTGCAGCTTTTTAGTCCTTTGTGCTCCCACGGGTCTACAGAGTCCCATCTGCCCAAAGGTCTTGAAGCTTGACAGGATGTTTTCGATTACTCAGTCTCCCAGGGCACTACTGGTCCGTAGGATTCGATTGGTCGGGGTAGGAGAGTTAAACAACATTTAAACAGAGTTCTCTCAAAAATGTCTAAAGGGATTGTAGGTAGATAACATCCAATCACTGTTTGCACTTATCTGAAATCTTCCCTCTTGGCTGCCCCCAGGTATTTACTGTGGAGAACATTGCATAGGAATGTCTGGAAAAAGCTTCTACAACTTGTTACAGCCTTCACATTTGTAGAAGCTTT 22 COX10-ND6-3′UTR*ATGGCCGCATCTCCGCACACTCTCTCCTCACGCCTCCTGACAGGTTGCGTAGGAGGCTCTGTCTGGTATCTTGAAAGAAGAACTATGATGTATGCTTTGTTTCTGTTGAGTGTGGGTTTAGTAATGGGGTTTGTGGGGTTTTCTTCTAAGCCTTCTCCTATTTATGGGGGTTTAGTATTGATTGTTAGCGGTGTGGTCGGGTGTGTTATTATTCTGAATTTTGGGGGAGGTTATATGGGTTTAATGGTTTTTTTAATTTATTTAGGGGGAATGATGGTTGTCTTTGGATATACTACAGCGATGGCTATTGAGGAGTATCCTGAGGCATGGGGGTCAGGGGTTGAGGTCTTGGTGAGTGTTTTAGTGGGGTTAGCGATGGAGGTAGGATTGGTGCTGTGGGTGAAAGAGTATGATGGGGTGGTGGTTGTGGTAAACTTTAATAGTGTAGGAAGCTGGATGATTTATGAAGGAGAGGGGTCAGGGTTGATTCGGGAGGATCCTATTGGTGCGGGGGCTTTGTATGATTATGGGCGTTGGTTAGTAGTAGTTACTGGTTGGACATTGTTTGTTGGTGTATATATTGTAATTGAGATTGCTCGGGGGAATTAGGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTCTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCA 23 COX10-opt_ND6-3′UTRATGGCCGCATCTCCGCACACTCTCTCCTCACGCCTCCTGACAGGTTGCGTAGGAGGCTCTGTCTGGTATCTTGAAAGAAGAACTATGATGTACGCCCTGTTCCTGCTGAGCGTGGGCCTGGTGATGGGCTTCGTGGGCTTCAGCAGCAAGCCCAGCCCCATCTACGGCGGCCTGGTGCTGATCGTGAGCGGCGTGGTGGGCTGCGTGATCATCCTGAACTTCGGCGGCGGCTACATGGGCCTGATGGTGTTCCTGATCTACCTGGGCGGCATGATGGTGGTGTTCGGCTACACCACCGCCATGGCCATCGAGGAGTACCCCGAGGCCTGGGGCAGCGGCGTGGAGGTGCTGGTGAGCGTGCTGGTGGGCCTGGCCATGGAGGTGGGCCTGGTGCTGTGGGTGAAGGAGTACGACGGCGTGGTGGTGGTGGTGAACTTCAACAGCGTGGGCAGCTGGATGATCTACGAGGGCGAGGGCAGCGGCCTGATCCGCGAGGACCCCATCGGCGCCGGCGCCCTGTACGACTACGGCCGCTGGCTGGTGGTGGTGACCGGCTGGACCCTGTTCGTGGGCGTGTACATCGTGATCGAGATCGCCCGCGGCAACTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCAGCCCCTGTCCTCCCTTCACCCCCATTGCGTATGAGCATTTCAGAACTCCAAGGAGTCACAGGCATCTTTATAGTTCACGTTAACATATAGACACTGTTGGAAGCAGTTCCTTCTAAAAGGGTAGCCCTGGACTTAATACCAGCCGGATACCTCTGGCCCCCACCCCATTACTGTACCTCTGGAGTCACTACTGTGGGTCGCCACTCCTCTGCTACACAGCACGGCTTTTTCAAGGCTGTATTGAGAAGGGAAGTTAGGAAGAAGGGTGTGCTGGGCTAACCAGCCCACAGAGCTCACATTCCTGTCCCTTGGGTGAAAAATACATGTCCATCCTGATATCTCCTGAATTCAGAAATTAGCCTCCACATGTGCAATGGCTTTAAGAGCCAGAAGCAGGGTTCTGGGAATTTTGCAAGTTACCTGTGGCCAGGTGTGGTCTCGGTTACCAAATACGGTTACCTGCAGCTTTTTAGTCCTTTGTGCTCCCACGGGTCTACAGAGTCCCATCTGCCCAAAGGTCTTGAAGCTTGACAGGATGTTTTCGATTACTCAGTCTCCCAGGGCACTACTGGTCCGTAGGATTCGATTGGTCGGGGTAGGAGAGTTAAACAACATTTAAACAGAGTTCTCTCAAAAATGTCTAAAGGGATTGTAGGTAGATAACATCCAATCACTGTTTGCACTTATCTGAAATCTTCCCTCTTGGCTGCCCCCAGGTATTTACTGTGGAGAACATTGCATAGGAATGTCTGGAAAAAGCTTCTACAACTTGTTACAGCCTTCACATTTGTAGAAGCTT T24 COX10-opt_ND6-3′UTR*ATGGCCGCATCTCCGCACACTCTCTCCTCACGCCTCCTGACAGGTTGCGTAGGAGGCTCTGTCTGGTATCTTGAAAGAAGAACTATGATGTACGCCCTGTTCCTGCTGAGCGTGGGCCTGGTGATGGGCTTCGTGGGCTTCAGCAGCAAGCCCAGCCCCATCTACGGCGGCCTGGTGCTGATCGTGAGCGGCGTGGTGGGCTGCGTGATCATCCTGAACTTCGGCGGCGGCTACATGGGCCTGATGGTGTTCCTGATCTACCTGGGCGGCATGATGGTGGTGTTCGGCTACACCACCGCCATGGCCATCGAGGAGTACCCCGAGGCCTGGGGCAGCGGCGTGGAGGTGCTGGTGAGCGTGCTGGTGGGCCTGGCCATGGAGGTGGGCCTGGTGCTGTGGGTGAAGGAGTACGACGGCGTGGTGGTGGTGGTGAACTTCAACAGCGTGGGCAGCTGGATGATCTACGAGGGCGAGGGCAGCGGCCTGATCCGCGAGGACCCCATCGGCGCCGGCGCCCTGTACGACTACGGCCGCTGGCTGGTGGTGGTGACCGGCTGGACCCTGTTCGTGGGCGTGTACATCGTGATCGAGATCGCCCGCGGCAACTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCA 25 COX10-ND1-3′UTRATGGCCGCATCTCCGCACACTCTCTCCTCACGCCTCCTGACAGGTTGCGTAGGAGGCTCTGTCTGGTATCTTGAAAGAAGAACTATGGCCAACCTCCTACTCCTCATTGTACCCATTCTAATCGCAATGGCATTCCTAATGCTTACCGAACGAAAAATTCTAGGCTATATGCAACTACGCAAAGGCCCCAACGTTGTAGGCCCCTACGGGCTACTACAACCCUCGCTGACGCCATAAAACTCTTCACCAAAGAGCCCCTAAAACCCGCCACATCTACCATCACCCTCTACATCACCGCCCCGACCTTAGCTCTCACCATCGCTCTTCTACTATGGACCCCCCTCCCCATGCCCAACCCCCTGGTCAACCTCAACCTAGGCCTCCTATTTATTCTAGCCACCTCTAGCCTAGCCGTTTACTCAATCCTCTGGTCAGGGTGGGCATCAAACTCAAACTACGCCCTGATCGGCGCACTGCGAGCAGTAGCCCAAACAATCTCATATGAAGTCACCCTAGCCATCATTCTACTATCAACATTACTAATGAGTGGCTCCTTTAACCTCTCCACCCTTATCACAACACAAGAACACCTCTGGTTACTCCTGCCATCATGGCCCTTGGCCATGATGTGGTTTATCTCCACACTAGCAGAGACCAACCGAACCCCCTTCGACCTTGCCGAAGGGGAGTCCGAACTAGTCTCAGGCTTCAACATCGAATACGCCGCAGGCCCCTTCGCCCTATTCTTCATGCCCGAATACACAAACATTATTATGATGAACACCCTCACCACTACAATCUCCTAGGAACAACATATCACGCACTCTCCCCTGAACTCTACACAACATATTTTGTCACCAAGACCCTACTTCTAACCTCCCTGTTCTTATGGATTCGAACAGCATACCCCCGATTCCGCTACGACCAACTCATGCACCTCCTATGGAAAAACTTCCTACCACTCACCCTAGCATTACTTATGTGGTATGTCTCCATGCCCATTACAATCTCCAGCATTCCCCCTCAAACCTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTCGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTCGACTGCCAGCCCCTGTCCTCCCTTCACCCCCATTCCGTATGAGCATTTCAGAACTCCAAGGAGTCACAGGCATCTTTATAGTTCACGTTAACATATAGACACTGTTGGAAGCAGTTCCTTCTAAAAGGGTAGCCCTGGACTTAATACCAGCCGGATACCTCTGGCCCCCACCCCATTACTGTACCTCTGGAGTCACTACTGTGGGTCGCCACTCCTCTGCTACACAGCACGGCTTTTTCAAGGCTGTATTGAGAAGGGAAGTTAGGAAGAAGGGTGTGCTGGGCTAACCAGCCCACAGAGCTCACATTCCTGTCCCTTGGGTGAAAAATACATGTCCATCCTGATATCTCCTGAATTCAGAAATTAGCCTCCACATGTGCAATGGCTTTAAGAGCCAGAAGCAGGGTTCTGGGAATTTTGCAAGTTACCTGTGGCCAGGTGTGGTCTCGGTTACCAAATACGGTTACCTGCAGCTTTTTAGTCCTTTGTGCTCCCACGGGTCTACAGAGTCCCATCTGCCCAAAGGTCTTGAAGCTTGACAGGATGTUTCGATTACTCAGTCTCCCAGGGCACTACTGGTCCGTAGGATTCGATTGGTCGGGCTAGGAGAGTTAAACAACATTTAAACAGAGTTCTCTCAAAAATGTCTAAAGGGATTGTAGGTAGATAACATCCAATCACTGTTTGCACTTATCTGAAATCTTCCCTCTTGGCTGCCCCCAGGTATTTACTGTGGAGAACATTGCATAGGAATGTCTGGAAAAAGCTTCTACAACTTGTTACAGCCTTCACATTTGTAGAAGCTTT 26COX10-ND1-3′UTR*ATGGCCGCATCTCCGCACACTCTCTCCTCACGCCTCCTGACAGGTTGCGTAGGAGGCTCTGTCTGGTATCTTGAAAGAAGAACTATGGCCAACCTCCTACTCCTCATTGTACCCATTCTAATCGCAATGGCATTCCTAATGCTTACCGAACGAAAAATTCTAGGCTATATGCAACTACGCAAAGGCCCCAACGTTGTAGGCCCCTACGGGCTACTACAACCCTTCGCTGACGCCATAAAACTCTTCACCAAAGAGCCCCTAAAACCCGCCACATCTACCATCACCCTCTACATCACCGCCCCGACCTTAGCTCTCACCATCGCTCTTCTACTATGGACCCCCCTCCCCATGCCCAACCCCCTGGTCAACCTCAACCTAGGCCTCCTATTTATTCTAGCCACCTCTAGCCTAGCCGTTTACTCAATCCTCTGGTCAGGGTGGGCATCAAACTCAAACTACGCCCTGATCGGCGCACTGCGAGCAGTAGCCCAAACAATCTCATATGAAGTCACCCTAGCCATCATTCTACTATCAACATTACTAATGAGTGGCTCCTTTAAACCTCTCCACCCTTATCACAACACAAGAACACCTCTGGTTACTCCTGCCATCATGGCCCTTGGCCATGATGTGGTTTATCTCCACACTAGCAGAGACCAACCGAACCCCCTTCGACCTTGCCGAAGGGGAGTCCGAACTAGTCTCAGGCTTCAACATCGAATACGCCGCAGGCCCCTTCGCCCTATTCTTCATGGCCCGAATACACAAACATTATTATGATGAACACCCTCACCACTACAATCCCTAGGAACAACATATCACGCACTCTCCCCTGTAACTCTACACAACATATTTTGTCACCAAGACCCTACTTCTAACCTCCCTGTTCTTATGGATTCGAACAGCATACCCCCGATTCCGCTACGACCAACTCATGCACCTCCTATGGAAAAACTTCCTACCACTCACCCTAGCATTACTTATGTGGTATGTCTCCATGCCCATTACAATCTCCAGCATTCCCCCTCAAACCTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATCGGGCTACACATACACAGCTTCCTCTTTTGCTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTCGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCA 27 COX10-opt_ND1-3′UTRATGGCCGCATCTCCGCACACTCTCTCCTCACGCCTCCTGACAGGTTGCGTAGGAGGCTCTGTCTGGTATCTTGAAAGAAAACTATGGCCAACCTGCTGCTGCTGATCGTGCCCATCCTGATCGCCATGGCCTTCCTGATGCTGACCGAGCGCAAGATCCTGGGCTACATGCAGCTGCGCAAGGGCCCCAACGTGGTGGGCCCCTACGGCCTGCTGCAGCCCTTCGCCGACGCCATCAAGCTGTTCACCAAGGAGCCCCTGAAGCCCGCCACCAGCACCATCACCCTGTACATCACCGCCCCCACCCTGGCCCTGACCATCGCCCTGCTGCTGTGGACCCCCCTGCCCATGCCCAACCCCCTGGTGAACCTGAACCTGGGCCTGCTGTTCATCCTGGCCACCAGCAGCCTGGCCGTGTACAGCATCCTGTGGAGCGGCTGGGCCAGCAACAGCAACTACGCCCTGATCGGCGCCCTGCGCGCCGTGGCCCAGACCATCAGCTACGAGGTGACCCTGGCCATCATCCTGCTGAGCACCCTGCTGATGAGCGGCAGCTTCAACCTGAGCACCCTGATCACCACCCAGGAGCACCTGTGGCTGCTGCTGCCCAGCTGGCCCCTGGCCATGATGTGGTTCATCAGCACCCTGGCCGAGACCAACCGCACCCCCTTCGACCTGGCCGAGGGCGAGAGCGAGCTGGTGAGCGGCTTCAACATCGAGTACGCCGCCGGCCCCTTCGCCCTGTTCTTCATGGCCGAGTACACCAACATCATCATGATGAACACCCTGACCACCACCATCTTCCTGGGCACCACCTACGACGCCCTGAGCCCCGAGCTCTACACCACCTACTTCGTGACCAAGACCCTGCTGCTGACCAGCCTGTTCCTGTGGATCCGCACCGCCTACCCCCGCTTCCGCTACGACCAGCTGATGCACCTGCTGTGGAAGAACTTCCTGCCCCTGACCCTGGCCCTGCTGATGTGGTACGTGAGCATGCCCATCACCATCAGCAGCATCCCCCCCCAGACCTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCAGCCCCTGTCCTCCCTTCACCCCCATTGCGTATGAGCATTTCAGAACTCCAAGGAGTCACAGGCATCTTTATAGTTCACGTTAACATATAGACACTGTTGGAAGCAGTTCCTTCTAAAAGGGTAGCCCTGGACTTAATACCAGCCGGATACCTCTGGCCCCCACCCCATTACTGTACCTCTGGAGTCACTACTGTGGGTCGCCACTCCTCTGCTACACAGCACGGCTTTTTCAAGGCTGTATTGAGAAGGGAAGTTAGGAAGAAGGGTGTGCTGGGCTAACCAGCCCACAGAGCTCACATTCCTGTCCCTTGGGTGAAAAATACATGTCCATCCTGATATCTCCTGAATTCAGAAATTAGCCTCCACATGTGCATGGCTTTAAGAGCCAGAAGCAGGGTTCTGGGAATTTTGCAAGTTACCTGTGGCCAGGTGTGGTCTCGGTTACCAAATACGGTTACCTGCAGCTTTTTAGTCCTTTGTGCTCCCACGGGTCTACAGAGTCCCATCTGCCCAAAGGTCTTGAAGCTTGACAGGATGTTTTCGATTACTCAGTCTCCCAGGGCACTACTGGTCCGTAGGATTCGATTGGTCGGGGTAGGAGAGTTAAACAACATTTAAACAGAGTTCTCTCAAAAATGTCTAAAGGGATTGTAGGTAGATAACATCCAATCACTGTTTGCACTTATCTGAAATCTTCCCTCTTGGCTGCCCCCAGGTATTTACTGTGGAGAACATTGCATAGGAATGTCTGGAAAAAGCTTCTACAACTTGTTACAGCCTTCACATTTGTAGAAGCTTT 28 COX10-opt_ND1-3′UTR*ATGGCCGCATCTCCGCACACTCTCTCCTCACGCCTCCTGACAGGTTGCGTAGGAGGCTCTGTCTGGTATCTTGAAAGAAGAACTATGGCCAACCTGCTGCTGCTGATCGTGCCCATCCTGATCGCCATGGCCTTCCTGATGCTGACCGAGCGCAAGATCCTGGGCTACATGCAGCTGCGCAAGGGCCCCAACGTGGTGGGCCCCTACGGCCTGCTGCAGCCCTTCGCCGACGCCATCAAGCTGTTCACCAAGGAGCCCCTGAAGCCCGCCACCAGCACCATCACCCTGTACATCACCGCCCCCACCCTCGCCCTGACCATCGCCCTGCTGCTGTGGACCCCCCTGCCCATGCCCAACCCCCTGGTGAACCTGAACCTGGGCCTGCTGTTCATCCTGGCCACCAGCAGCCTGGCCGTGTACAGCATCCTGTGGAGCGGCTGGGCCAGCAACAGCAACTACGCCCTGATCGGCGCCCTGCGCGCCGTGGCCCAGACCATCAGCTACGAGGTGACCCTGGCCATCATCCTGCTGAGCACCCTCCTGATGAGCGGCAGCTTCAACCTGAGCACCCTGATCACCACCCAGGAGCACCTGTGGCTGCTGCTGCCCAGCTGGCCCCTGGCCATGATGTGGTTCATCAGCACCCTGGCCGAGACCAACCGCACCCCCTTCGACCTGGCCGAGGGCGAGAGCGAGCTGGTGAGCGGCTTCAACATCGAGTACGCCGCCGGCCCCTTCGCCCTGTTCTTCATGGCCGAGTACACCAACATCATCATGATGAACACCCTGACCACCACCATCTTCCTGGGCACCACCTACGACGCCCTGAGCCCCGAGCTGTACACCACCTACTTCGTGACCAAGACCCTGCTGCTGACCAGCCTGTTCCTGTGGATCCGCACCGCCTACCCCCGCTTCCGCTACGACCAGCTGATGCACCTGCTGTGGAAGAACTTCCTGCCCCTGACCCTGGCCCTGCTGATGTGGTACGTGAGCATGCCCATCACCATCAGCAGCATCCCCCCCCAGACCTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCA 29 opt_COX10-ND4-3′UTRATGGCCGCCTCTCCACACACACTGAGTAGCAGACTGCTGACCGGCTGTGTTGGCGGCTCTGTGTGGTATCTGGAACGGCGGACAATGCTAAAACTAATCGTCCCAACAATTATGTTACTACCACTGACATGGCTTTCCAAAAAACACATGATTTGGATCAACACAACCACCCACAGCCTAATTATTAGCATCATCCCTCTACTATTTTTTAACCAAATCAACAACAACCTATTTAGCTGTTCCCCAACCTTTTCCTCCGACCCCCTAACAACCCCCCTCCTAATGCTAACTACCTGGCTCCTACCCCTCACAATCATGGCAAGCCAACGCCACTTATCCAGTGAACCACTATCACGAAAAAAACTCTACCTCTCTATGCTAATCTCCCTACAAATCTCCTTAATTATGACATTCACAGCCACAGAACTAATCATGTTTTATATCTTCTTCGAAACCACACTTATCCCCACCTTGGCTATCATCACCCGATGGGGCAACCAGCCAGAACGCCTGAACGCAGGCACATACTTCCTATTCTACACCCTAGTAGGCTCCCTTCCCCTACTCATCGCACTAATTTACACTCACAACACCCTAGGCTCACTAAACATTCTACTACTCACTCTCACTGCCCAAGAACTATCAAACTCCTGGGCCAACAACTTAATGTGGCTAGCTTACACAATGGCTTTTATGGTAAAGATGCCTCTTTACGGACTCCACTTATGGCTCCCTAAAGCCCATGTCGAAGCCCCCATCGCTGGGTCAATGGTACTTGCCGCAGTACTCTTAAAACTAGGCGGCTATGGTATGATGCGCCTCACACTCATTCTCAACCCCCTGACAAAACACATGGCCTACCCCTTCCTTGTACTATCCCTATGGGGCATGATTATGACAAGCTCCATCTGCCTACGACAAACAGACCTAAAATCGCTCATTGCATACTCTTCAATCAGCCACATGGCCCTCGTAGTAACAGCCATTCTCATCCAAACCCCCTGGAGCTTCACCGGCGCAGTCATTCTCATGATCGCCCACGGGCTTACATCCTCATTACTATTCTGCCTAGCAAACTCAAACTACGAACGCACTCACAGTCGCATCATGATCCTCTCTCAAGGACTTCAAACTCTACTCCCACTAATGGCTTTTTGGTGGCTTCTAGCAAGCCTCGCTAACCTCGCCTTACCCCCCACTATTAACCTACTGGGAGAACTCTCTGTGCTAGTAACCACGTTCTCCTGGTCAAATATCACTCTCCTACTTACAGGACTCAACATGCTAGTCACAGCCCTATACTCCCTCTACATGTTTACCACAACACAATGGGGCTCACTCACCCACCACATTAACAACATGAAACCCTCATTCACACGAGAAAACACCCTCATGTTCATGCACCTATCCCCCATTCTCCTCCTATCCCTCAACCCCGACATCATTACCGGGTTTTCCTCTTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCAGCCCCTGTCCTCCCTTCACCCCCATTGCGTATGAGCATTTCAGAACTCCAAGGAGTCACAGGCATCTTTATAGTTCACGTTAACATATAGACACTGTTGGAAGCAGTTCCTTCTAAAAGGGTAGCCCTGGACTTAATACCAGCCGGATACCTCTGGCCCCCACCCCATTACTGTACTCTGGAGTCACTACTGTGGGTCGCCACTCCTCTGCTACACAGCACGGCTTTTTCAAGGCTGTATTGAGAAGGGAAGTTAGGAAGAAGGGTGTGCTGGGCTAACCAGCCCACAGAGCTCACATTCCTGTCCCTTGGGTGAAAAATACATGTCCATCCTGATATCTCCTGAATTCAGAAATTAGCCTCCACATGTGCAATGGCTTTAAGAGCCAGAAGCAGGGTTCTGGGAATTTTGCAAGTTACCTGTGGCCAGGTGTGGTCTCGGTTACCAAATACGGTTACCTGCAGCTTTTTAGTCCTTTGTGCTCCCACGGGTCTACAGAGTCCCATCTGCCCAAAGGTCTTGAAGCTTGACAGGATGTTTTCGATTACTCAGTCTCCCAGGGCACTACTGGTCCGTAGGATTCGATTGGTCGGGGTAGGAGAGTTAAACAACATTTAAACAGAGTTCTCTCAAAAATGTCTAAAGGGATTGTAGGTAGATAACATCCAATCACTGTTTGCACTTATCTGAAATCTTCCCTCTTGGCTGCCCCCAGGTATTTACTGTGGAGAACATTGCATAGGAATGTCTGGAAAAAGCTTCTACAACTTGTTACAGCCTTCACATTTGTAGAAGCTTT 30 opt_COX10-ND4-3′UTR*ATGGCCGCCTCTCCACACACACTGAGTAGCAGACTGCTGACCGGCTGTGTTGGCGGCTCTGTGTGGTATCTGGAACGGCGGACAATGCTAAAACTAATCGTCCCAACAATTATGTTACTACCACTGACATGGCTTTCCAAAAAACACATGATTTGGATCAACACCCACCCACAGCCTAATTATTAGCATCATCCCTCTACTATTTTTTAACCAAATCAACAACAACCTATTTAGCTGTTCCCCAACCTTTTCCTCCGACCCCCTAACAACCCCCCTCCTAATGCTAACTACCTGGCTCCTACCCCTCACAATCATGGCAAGCCAACGCCACTTATCCAGTGAACCACTATCACGAAAAAAACTCTACCTCTCTATGCTAATCTCCCTACAAATCTCCTTAATTATGACATTCACAGCCACAGAACTAATCATGTTTTATATCTTCTTCGAAACCACACTTATCCCCACCTTGGCTATCATCACCCGATGGGGCAACCAGCCAGAACGCCTGAACGCAGGCACATACTTCCTATTCTACACCCTAGTAGGCTCCCTTCCCCTACTCATCGCACTAATTTACACTCACAACACCCTAGGCTCACTAAACATTCTACTACTCACTCTCACTGCCCAAGAACTATCAAACTCCTGGGCCAACAACTTAATGTGGCTAGCTTACACAATGGCTTTTATGGTAAAGATGCCTCTTTACGGACTCCACTTATGGCTCCCTAAGCCCATGTCGAAGCCCCCATCGCTGGGTCAATGGTACTTGCCGCAGTACTCTTAAAACTAGGCGGCTATGGTATGATGCGCCTCACACTCATTCTCAACCCCCTGACAAAACACATGGCCTACCCCTTCCTTGTACTATCCCTATGGGGCATGATTATGACAAGCTCCATCTGCCTACGACAAACAGACCTAAAATCGCTCATTGCATACTCTTCAATCAGCCACATGGCCCTCGTAGTAACAGCCATTCTCATCCAAACCCCCTGGAGCTTCACCGGCGCAGTCATTCTCATGATCGCCCACGGGCTTACATCCTCATTACTATTCTGCCTAGCAAACTCAAACTACGAACGCACTCACAGTCGCATCATGATCCTCTCTCAAGGACTTCAAACTCTACTCCCACTAATGGCTTTTTGGTGGCTTCTAGCAAGCCTCGCTAACCTCGCCTTACCCCCCACTATTAACCTACTGGGAGAACTCTCTGTGCTAGTAACCACGTTCTCCTGGTCAAATATCACTCTCCTACTTACAGGACTCAACATGCTAGTCACAGCCCTATACTCCCTCTACATGTTTACCACAACACAATGGGGCTCACTCACCCACCACATTAACAACATGAAACCCTCATTCACACGAGAAAACACCCTCATGTTCATGCACCTATCCCCCATTCTCCTCCTATCCCTCAACCCCGACATCATTACCGGGTTTTCCTCTTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCA 31 opt_COX10-opt_ND4-3′UTRATGGCCGCCTCTCCACACACACTGAGTAGCAGACTGCTGACCGGCTGTGTTGGCGGCTCTGTGTGGTATCTGGAACGGCGGACAATGCTGAAGCTGATCGTGCCCACCATCATGCTGCTGCCTCTGACCTGGCTGAGCAAGAAACACATGATCTGGATCAACACCACCACGCACAGCCTGATCATCAGCATCATCCCTCTGCTGTTCTTCAACCAGATCAACAACAACCTGTTCAGCTGCAGCCCCACCTTCAGCAGCGACCCTCTGACAACACCTCTGCTGATGCTGACCACCTGGCTGCTGCCCCTCACAATCATGGCCTCTCAGAGACACCTGAGCAGCGAGCCCCTGAGCCGGAAGAAACTGTACCTGAGCATGCTGATCTCCCTGCAGATCTCTCTGATCATGACCTTCACCGCCACCGAGCTGATCATGTTCTACATCTTTTTCGAGACAACGCTGATCCCCACACTGGCCATCATCACCAGATGGGGCAACCAGCCTGAGAGACTGAACGCCGGCACCTACTTTCTGTTCTACACCCTCGTGGGCAGCCTGCCACTGCTGATTGCCCTGATCTACACCCACAACACCCTGGGCTCCCTGAACATCCTGCTGCTGACACTGACAGCCCAAGAGCTGAGCAACAGCTGGGCCAACAATCTGATGTGGCTGGCCTACACAATGGCCTTCATGGTCAAGATGCCCCTGTACGGCCTGCACCTGTGGCTGCCTAAAGCTCATGTGGAAGCCCCTATCGCCGGCTCTATGGTGCTGGCTGCAGTGCTGCTGAAACTCGGCGGCTACGGCATGATGCGGCTGACCCTGATTCTGAATCCCCTGACCAAGCACATGGCCTATCCATTTCTGGTGCTGAGCCTGTGGGGCATGATTATGACCAGCAGCATCTGCCTGCGGCAGACCGATCTGAAGTCCCTGATCGCCTACAGCTCCATCAGCCACATGGCCCTGGTGGTCACCGCCATCCTGATTCAGACCCCTTGGAGCTTTACAGGCGCCGTGATCCTGATGATTGCCCACGGCCTGACAAGCAGCCTGCTGTTTTGTCTGGCCAACAGCAACTACGAGCGGACCCACAGCAGAATCATGATCCTGTCTCAGGGCCTGCAGACCCTCCTGCCTCTTATGGCTTTTTGGTGGCTGCTGGCCTCTCTGGCCAATCTGGCACTGCCTCCTACCATCAATCTGCTGGGCGAGCTGAGCGTGCTGGTCACCACATTCAGCTGGTCCAATATCACCCTGCTGCTCACCGGCCTGAACATGCTGGTTACAGCCCTGTACTCCCTGTACATGTTCACCACCACACAGTGGGGAAGCCTGACACACCACATCAACAATATGAAGCCCAGCTTCACCCGCGAGAACACCCTGATGTTCATGCATCTGAGCCCCATTCTGCTGCTGFCCCTGAATCCTGATATCATCACCGGCTTCTCCAGCTGAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCAGCCCCTGTCCTCCCTTCACCCCCATTGCGTATGAGCATTTCAGAACTCCAAGGAGTCACAGGCATCTTTATAGTTCACGTTAACATATAGACACTGTTGGAAGCAGTTCCTTCTAAAAGGGTAGCCCTGGACTTAATACCAGCCGGATACCTCTGGCCCCCACCCCATTACTGTACCTCTGGAGTCACTACTGTGGGTCGCCACTCCTCTGCTACACAGCACGGCTTTTTCAGGCTGTATTGAGAAGGGAAGTTAGGAAGAAAGGGTGTGCTGGGCTAACCAGCCCACAGAGCTCACATTCCTGTCCCTTGGGTGAAAAATACATGTCCATCCTGATATCTCCTGAATTCAGAAATTAGCCTCCACATGTGCAATGGCTTTAAGAGCCAGAAGCAGGGTTCTGGGAATTTTGCAAGTTACCTGTGGCCAGGTGTGGTCTCGGTTACCAAATACGGTTACCTGCAGCTTTTTAGTCCTTTGTGCTCCCACGGGTCTACAGAGTCCCATCTGCCCAAAGGTCTTGAAGCTTGACAGGATGTTTTCGATTACTCAGTCTCCCAGGGCACTACTGGTCCGTAGGATTCGATTGGTCGGGGTAGGAGAGTTAAACAACATTTAAACAGAGTTCTCTCAAAAATGTCTAAAGGGATTGTAGGTAGATAACATCCAATCACTGTTTGCACTTATCTGAAATCTTCCCTCTTGGCTGCCCCCAGGTATTTACTGTGGAGAACATTGCATAGGAATGTCTGGAAAAAGCTTCTACAACTTGTTACAGCCTTCACATTTGTAGAAGCTTT 32 opt_COX10-opt_ND4-3′UTR* ATGGCCGCCTCTCCACACACACTGAGTAGCAGACTGCTGACCGGCTGTGTTGGCGGCTCTGTGTGGTATCTGGAACGGCGGACAATGCTGAAGCTGATCGTGCCCACCATCATGCTGCTGCCTCTGACCTGGCTGAGCAAGAAACACATGATCTGGATCAACACCACCACGCACAGCCTGATCATCAGCATCATCCCTCTGCTGTTCTTCAACCAGATCAACAACAACCTGTTCAGCTGCAGCCCCACCTTCAGCAGCGACCCTCTGACAACACCTCTGCTGATGCTGACCACCTGGCTGCTGCCCCTCACAATCATGGCCTCTCAGAGACACCTGAGCAGCGAGCCCCTGAGCCGGAAGAAACTGTACCTGAGCATGCTGATCTCCCTGCAGATCTCTCTGATCATGACCTTCACCGCCACCGAGCTGATCATGTTCTACATCTTTTTCGAGACAACGCTGATCCCCACACTGGCCATCATCACCAGATGGGGCAACCAGCCTGAGAGACTGAACGCCGGCACCTACTTTCTGTTCTACACCCTCGTGGGCAGCCTGCCACTGCTGATTGCCCTGATCTACACCCACAACACCCTGGGCTCCCTGAACATCCTGCTGCTGACACTGACAGCCCAAGAGCTGAGCAACAGGTGGGCCAACAATCTGATGTGGCTGGCCTACACAATGGCCTTCATGGTCAAGATGCCCCTGTACGGCCTGCACCTGTGGCTGCCTAAAGCTCATGTGGAAGCCCCTATCGCCGGCTCTATGGTGCTGGCTGCAGTGCTGCTGAAACTCGGCGGCTACGGCATGATGCGGCTGACCCTGATTCTGAATCCCCTGACCAAGCACATGGCCTATCCATTTCTGGTGCTGAGCCTGTGGGGCATGATTATGACCAGCAGCATCTGCCTGCGGCAGACCGATCTGAAGTCCCTGATCGCCTACAGCTCCATCAGCCACATGGCCCTGGTGGTCACCGCCATCCTGATTCAGACCCcrTGGAGCTTTACAGGCGCCGTGATCCTGATGATTGCCCACGGCCTGACAAGCAGCCTGCTGTTTTGTCTGGCCAACAGCAACTACGAGCGGACCCACAGCAGAATCATGATCCTGTCTCAGGGCCTGCAGACCCTCCTGCCTCTTATGGCTTTTTGGTGGCTGCTGGCCTCTCTGGCCAATCTGGCACTGCCTCCTACCATCAATCTGCTGGGCGAGCTGAGCGTGCTGGTCACCACATTCAGGTGGTCCAATATCACCCTGCTGCTCACCGGCCTGAACATGCTGGTTACAGCCCTGTACTCCCTGTACATGTTCACCACCACACAGTGGGGAAGCCTGACACACCACATCAACAATATGAAGCCCAGCTTCACCCGCGAGAACACCCTGATGTTCATGCATCTGAGCCCCATTCTGCTGCTGTCCCTGAATCCTGATATCATCACCGGCTTCTCCAGCTGAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGGTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCA 33opt_COX10-opt_ND4*-3′UTRATGGCCGCCTCTCCACACACACTGAGTAGCAGACTGCTGACCGGCTGTGTTGGCGGCTCTGTGTGGTATCTGGAACGGCGGACAATGCTGAAGCTGATCGTGCCCACCATCATGCTGCTGCCCCTGACCTGGCTGAGCAAGAAGCACATGATCTGGATCAACACCACCACCCACAGCCTGATCATCAGCATCATCCCCCTGCTGTTCTTCAACCAGATCAACAACAACCTGTTCAGCTGCAGCCCCACCTTCAGCAGCGACCCCCTGACCACCCCCCTGCTGATGCTGACCACCTGGCTGCTGCCCCTGACCATCATGGCCAGCCAGCGCCACCTGAGCAGCGAGCCCCTGAGCCGCAAGAAGCTGTACCTGAGCATGCTGATCAGCCTGCAGATCAGCCTGATCATGACCTTCACCGCCACCGAGCTGATCATGTTCTACATCTTCTTCGAGACCACCCTGATCCCCACCCTGGCCATCATCACCCGCTGGGGCAACCAGCCCGAGCGCCTGAACGCCGGCACCTACTTCCTGTTCTACACCCTGGTGGGCAGCCTGCCCCTGCTGATCGCCCTGATCTACACCCACAACACCCTGGGCAGCCTGAACATCCTGCTGCTGACCCTGACCGCCCAGGAGCTGAGCAACAGCTGGGCCAACAACCTGATGTGGCTGGCCTACACCATGGCCTTCATGGTGAAGATGCCCCTGTACGGCCTGCACCTGTGGCTGCCCAAGGCCCACGTGGAGGCCCCCATCGCCGGCAGCATGGTGCTGGCCGCCGTGCTGCTGAAGCTGGGCGGCTACGGCATGATGCGCCTGACCCTGATCCTGAACCCCCTGACCAAGCACATGGCCTACCCCTTCCTGGTGCTGAGCCTGTGGGGCATGATCATGACCAGCAGCATCTGCCTGCGCCAGACCGACCTGAAGAGCCTGATCGCCTACAGCAGCATCAGCCACATGGCCCTGGTGGTGACCGCCATCCTGATCCAGACCCCCTGGAGCTTCACCGGCGCCGTGATCCTGATGATCGCCCACGGCCTGACCAGCAGCCTGCTGTTCTGCCTGGCCAACAGCAACTACGAGCGCACCCACAGCCGCATCATGATCCTGAGCCAGGGCCTGCAGACCCTGCTGCCCCTGATGGCCTTCTGGTGGCTGCTGGCCAGCCTGGCCAACCTGGCCCTGCCCCCCACCATCAACCTGCTGGGCGAGCTGAGCGTGCTGGTGACCACCTTCAGCTGGAGCAACATCACCCTGCTGCTGACCGGCCTGAACATGCTGGTGACCGCCCTGTACAGCCTGTACATGTTCACCACCACCCAGTGGGGCAGCCTGACCCACCACATCAACAACATGAAGCCCAGCTTCACCCGCGAGAACACCCTGATGTTCATGCACCTGAGCCCCATCCTGCTGCTGAGCCTGAACCCCGACATCATCACCGGCTTCAGCAGCTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCAGCCCCTGTCCTCCCTTCACCCCCATTGCGTATGAGCATTTCAGAACTCCAAGGAGTCACAGGCATCTTTATAGTTCACGTTAACATATAGACACTGTTGGAAGCAGTTCCTTCTAAAAGGGTAGCCCTGGACTTAATACCAGCCGGATACCTCTGGCCCCCACCCCATTACTGTACCTCTGGAGTCACTACTGTGGGTCGCCACTCCTCTGCTACACAGCACGGCTTTTTCAAGGCTGTATTGAGAAGGGAAGTTAGGAAGAAGGGTGTGCTGGGCTAACCAGCCCACAGAGCTCACATTCCTGTCCCTTGGGTGAAAAATACATGTCCATCCTGATATCTCCTGAATTCAGAAATTAGCCTCCACATGTGCAATGGCTTTAAGAGCCAGAAGCAGGGTTCTGGGAATTTTGCAAGTTACCTGTGGCCAGGTGTGGTCTCGGTTACCAAATACGGTTACCTGCAGCTTTTTAGTCCTTTGTGCTCCCACGGGTCTACAGAGTCCCATCTGCCCAAAGGTCTTGAAGCTTGACAGGATGTTTTCGATTACTCAGTCTCCCAGGGCACTACTGGTCCGTAGGATTCGATTGGTCGGGGTAGGAGAGTTAAACAACATTTAAACAGAGTTCTCTCAAAAATGTCTAAAGGGATTGTAGGTAGATAACATCCAATCACTGTTTGCACTTATCTGAAATCTTCCCTCTTGGCTGCCCCCAGGTATTTACTGTGGAGAACATTGCATAGGAATGTCTGGAAAAAGCTTCTACAACTTGTTACAGCCTTCACATTTGTAGAAGCTTT 34opt_COX10-opt_ND4*-3′UTR*ATGGCCGCCTCTCCACACACACTGAGTAGCAGACTGCTGACCGGCTGTGTTGGCGGCTCTGTGTGGTATCTGGAACGGCGGACAATGCTGAAGCTGATCGTGCCCACCATCATGCTGCTGCCCCTGACCTGGCTGAGCAAGAAGCACATGATCTGGATCAACACCACCACCCACAGCCTGATCATCAGCATCATCCCCCTGCTGTTCTTCAACCAGATCAACAACAACCTGTTCAGCTGCAGCCCCACCTTCAGCAGCGACCCCCTGACCACCCCCCTGCTGATGCTGACCACCTGGCTGCTGCCCCTGACCATCATGGCCAGCCAGCGCCACCTGAGCAGCGAGCCCCTGAGCCGCAAGAAGCTGTACCTGAGCATGCTGATCAGCCTGCAGATCAGCCTGATCATGACCTTCACCGCCACCGAGCTGATCATGTTCTACATCTTCTTCGAGACCACCCTGATCCCCACCCTGGCCATCATCACCCGCTGGGGCAACCAGCCCGAGCGCCTGAACGCCGGCACCTACTTCCTGTTCTACACCCTGGTGGGCAGCCTGCCCCTGCTGATCGCCCTGATCTACACCCACAACACCCTGGGCAGCCTGAACATCCTGCTGCTGACCCTGACCGCCCAGGAGCTGAGCAACAGCTGGGCCAACAACCTGATGTGGCTGGCCTACACCATGGCCTTCATGGTGAAGATGCCCCTGTACGGCCTGCACCTGTGGCTGCCCAAGGCCCACGTGGAGGCCCCCATCGCCGGCAGCATGGTGCTGGCCGCCGTGCTGCTGAAGCTGGGCGGCTACGGCATGATGCGCCTGACCCTGATCCTGAACCCCCTGACCAAGCACATGGCCTACCCCTTCCTGGTGCTGAGCCTGTGGGGCATGATCATGACCAGCAGCATCTGCCTGCGCCAGACCGACCTGAAGAGCCTGATCGCCTACAGCAGCATCAGCCACATGGCCCTGGTGGTGACCGCCATCCTGATCCAGACCCCCTGGAGCTTCACCGGCGCCGTGATCCTGATGATCGCCCACGGCCTGACCAGCAGCCTGCTGTTCTGCCTGGCCAACAGCAACTACGAGCGCACCCACAGCCGCATCATGATCCTGAGC AGGGCCTGCAGACCCTGCTGCCCCTGATGGCCTTCTGGTGGCTGCTGGCCAGCCTGGCCAACCTGGCCCTGCCCCCCACCATCAACCTGCTGGGCGAGCTGAGCGTGGTGGTGACCACCTTCAGCTGGAGCAACATCACCCTGCTGCTGACCGGCCTGAACATGCTGGTGACCGCCCTGTACAGCCTGTACATGTTCACCACCACCCAGTGGGGCAGCCTGACCCACCACATCAACAACATGAAGCCCAGCTTCACCCGCGAGAACACCCTGATGTTCATGCACCTGAGCCCCATCCTGCTGCTGAGCCTGAACCCCGACATCATCACCGGCTTCAGCAGCTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCA 35opt_COX10-ND6-3′UTRATGGCCGCCTCTCCACACACACTGAGTAGCAGACTGCTGACCGGCTGTGTTGGCGGCTCTGTGTGGTATCTGGAACGGCGGACAATGATGTATGCTTTGTTTCTGTTGAGTGTGGGTTTAGTAATGGGGTTTGTGGGGTTTTCTTCTAAGCCTTCTCCTATTTATGGGGGTTTAGTATTGATTGTTAGCGGTGTGGTCGGGTGTGTTATTATTCTGAATTTTGGGGGAGGTTATATGGGTTTAATGGTTTTTTTAATTTATTTAGGGGGAATGATGGTTGTCTTTGGATATACTACAGCGATGGCTATTGAGGAGTATCCTGAGGCATGGGGGTCAGGGGTTGAGGTCTTGGTGAGTGTTTTAGTGGGGTTAGCGATGGAGGTAGGATTGGTGCTGTGGGTGAAAGAGTATGATGGGGTGGTGGTTGTGGTAAACTTTAATAGTGTAGGAAGCTGGATGATTTATGAAGGAGAGGGGTCAGGGTTGATTCGGGAGGATCCTATTGGTGCGGGGGCTTTGTATGATTATGGGCGTTGGTTAGTAGTAGTTACTGGTTGGACATTGTTTGTTGGTGTATATATTGTAATTGAGATTGCTCGGGGGAATTAGGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCAGCCCCTGTCCTCCCTTCACCCCCATTGCGTATGAGCATTTCAGAACTCCAAGGAGTCACAGGCATCTTTATAGTTCACGTTAACATATAGACACTGTTGGAAGCAGTTCCTTCTAAAAGGGTAGCCCTGGACTTAATACCAGCCGGATACCTCTGGCCCCCACCCCATTACTGTACCTCTGGAGTCACTACTGTGGGTCGCCACTCCTCTGCTACACAGCACGGCTTTTTCAAGGCTGTATTGAGAAGGGAAGTTAGGAAGAAGGGTGTGCTGGGCTAACCAGCCCACAGAGCTCACATTCCTGTCCCTTGGGTGAAAAATACATGTCCATCCTGATATCTCCTGAATTCAGAAATTAGCCTCCACATGTGCAATGGCTTTAAGAGCCAGAAGCAGGGTTCTGGGAATTTTGCAAGTTACCTGTGGCCAGGTGTGGTCTCGGTTACCAAATACGGTTACCTGCAGCTTTTTAGTCCTTTGTGCTCCCACGGGTCTACAGAGTCCCATCTGCCCAAAGGTCTTGAAGCTTGACAGGATGTTTTCGATTACTCAGTCTCCCAGGGCACTACTGGTCCGTAGGATTCGATTGGTCGGGGTAGGAGAGTTAAACAACATTTAAACAGAGTTCTCTCAAAAATGTCTAAAGGGATTGTAGGTAGATAACATCCAATCACTGTTTGCACTTATCTGAAATCTTCCCTCTTGGCTGCCCCCAGGTATTTACTGTGGAGAACATTGCATAGGAATGTCTGGAAAAAGCTTCTACAACTTGTTACAGCCTTCACATTTGTAGAAGCTTT 36opt_COX10-ND6-3′UTR*ATGGCCGCCTCTCCACACACACTGAGTAGCAGACTGCTGACCGGCTGTGTTGGCGGCTCTGTGTGGTATCTGGAACGGCGGACAATGATGTATGCTTTGTTTCTGTTGAGTGTGGGTTTAGTAATGGGGTTTGTGGGGTTTTCTTCTAAGCCTTCTCCTATTTATGGGGGTTTAGTATTGATTGTTAGCGGTGTGGTCGGGTGTGTTATTATTCTGAATTTTGGGGGAGGTTATATGGGTTTAATGGTTTTTTTAATTTATTTAGGGGGAATGATGGTTGTCTTTGGATATACTACAGCGATGGCTATTGAGGAGTATCCTGAGGCATGGGGGTCAGGGGTTGAGGTCTTGGTGAGTGTTTTAGTGGGGTTAGCGATGGAGGTAGGATTGGTGCTGTGGGTGAAAGAGTATGATGGGGTGGTGGTTGTGGTAAACTTTAATAGTGTAGGAAGCTGGATGATTTATGAAGGAGAGGGGTCAGGGTTGATTCGGGAGGATCCTATTGGTGCGGGGGCTTTGTATGATTATGGGCGTTGGTTAGTAGTAGTTACTGGTTGGACATTGTTTGTTGGTGTATATATTGTAATTGAGATTGCTCGGGGGAATTAGGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCA 37 opt_COX10-opt_NDS-3′UTRATGGCCGCCTCTCCACACACACTGAGTAGCAGACTGCTGACCGGCTGTGTTGGCGGCTCTGTGTGGTATCTGGAACGGCGGACAATGATGTACGCCCTGTTCCTGCTGAGCGTGGGCCTGGTGATGGGCTTCGTGGGCTTCAGCAGCAAGCCCAGCCCCATCTACGGCGGCCTGGTGCTGATCGTGAGCGGCGTGGTGGGCTGCGTGATCATCCTGAACTTCGGCGGCGGCTACATGGGCCTGATGGTGTTCCTGATCTACCTGGGCGGCATGATGGTGGTGTTCGGCTACACCACCGCCATGGCCATCGAGGAGTACCCCGAGGCCTGGGGCAGCGGCGTGGAGGTGCTGGTGAGCGTGCTGGTGGGCCTGGCCATGGAGGTGGGCCTGGTGCTGTGGGTGAAGGAGTACGACGGCGTGGTGGTGGTGGTGAACTTCAACAGCGTGGGCAGCTGGATGATCTACGAGGGCGAGGGCAGCGGCCTGATCCGCGAGGACCCCATCGGCGCCGGCGCCCTGTACGACTACGGCCGCTGGCTGGTGGTGGTGACCGGCTGGACCCTGTTCGTGGGCGTGTACATCGTGATCGAGATCGCCCGCGGCAACTAAGAGCACTGGGACGCCCACCGCCCCTTTCccTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCAGCCCCTGTCCTCCCTTCACCCCCATTGCGTATGAGCATTTCAGAACTCCAAGGAGTCACAGGCATCTTTATAGTTCACGTTAAACATATAGACACTGTTGGAAGCAGTTCCTTCTAAAAGGGTAGCCCTGGACTTAATACCAGCCGGATACCTCTGGCCCCCACCCCATTACTGTACCTCTGGAGTCACTACTGTGGGTCGCCACTCCTCTGCTACACAGCACGGCTTTTTCAAGGCTGTATTGAGAAGGGAAGTTAGGAAGAAGGGTGTGCTGGGCTAACCAGCCCACAGAGCTCACATTCCTGTCCCTTGGGTGAAAAATACATGTCCATCCTGATATCTCCTGAAATTCAGAAATTAGCCTCCACATGTGCAATGGCTTTAAGAGCCAGAAGCAGGGTTCTGGGAATTTTGCAAGTTACCTGTGGCCAGGTGTGGTCTCGGTTACCAAATACGGTTACCTGCAGGTTTTTAGTCCTTTGTGCTCCCACGGGTCTACAGAGTCCCATCTGCCCAAAGGTCTTGAAGCTTGACAGGATGTTTTCGATTACTCAGTCTCCCAGGGCACTACTGGTCCGTAGGATTCGATTGGTCGGGGTAGGAGAGTTAAACAACATTTAAACAGAGTTCTCTCAAAAATGTCTAAAGGGATTGTAGGTAGATAACATCCAATCACTGTTTGCACTTATCTGAAATCTTCCCTCTTGGCTGCCCCCAGGTATTTACTGTGGAGAACATTGCATAGGAATGTCTGGAAAAAGCTTCTACAACTTGTTACAGCCTTCACATTTGTAGAAGCTT T38 opt_COX10-opt_ND6-3′UTR*ATGGCCGCCTCTCCACACACACTGAGTAGCAGACTGCTGACCGGCTGTGTTGGCGGCTCTGTGTGGTATCTGGAACGGCGGACAATGATGTACGCCCTGTTCCTGCTGAGCGTGGGCCTGGTGATGGGCTTCGTGGGCTTCAGCAGCAAGCCCAGCCCCATCTACGGCGGCCTGGTGCTGATCGTGAGCGGCGTGGTGGGCTGCGTGATCATCCTGAACTTCGGCGGCGGCTACATGGGCCTGATGGTGTTCCTGATCTACCTGGGCGGCATGATGGTGGTGTTCGGCTACACCACCGCCATGGCCATCGAGGAGTACCCCGAGGCCTGGGGCAGCGGCGTGGAGGTGCTGGTGAGCGTGCTGGTGGGCCTGGCCATGGAGGTGGGCCTGGTGCTGTGGGTGAAGGAGTACGACGGCGTGGTGGTGGTGGTGAACTTCAACAGCGTGGGCAGCTGGATGATCTACGAGGGCGAGGGCAGCGGCCTGATCCGCGAGGACCCCATCGGCGCCGGCGCCCTGTACGACTACGGCCGCTGGCTGGTGGTGGTGACCGGCTGGACCCTGTTCGTGGGCGTGTACATCGTGATCGAGATCGCCCGCGGCAACTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAATTGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCA 39 opt_COX10-ND1-3′UTRATGGCCGCCTCTCCACACACACTGAGTAGCAGACTGCTGACCGGCTGTGTTGGCGGCTCTGTGTGGTATCTGGAACGGCGGACAATGGCCAACCTCCTACTCCTCATTGTACCCATTCTAATCGCAATGGCATTCCTAATGCTTACCGAACGAAAAATTCTAGGCTATATGCAACTACGCAAAGGCCCCAACGTTGTAGGCCCCTACGGGCTACTACAACCCTTCGCTGACGCCATAAACTCTTCACCAAAGAGCCCCTAAAACCCGCCACATCTACCATCACCCTCTACATCACCGCCCCGACCTTAGCTCTCACCATCGCTCTTCTACTATGGACCCCCCTCCCCATGCCCAACCCCCTGGTCAACCTCAACCTAGGCCTCCTATTTATTCTAGCCACCTCTAGCCTAGCCGTTTACTCAATCCTCTGGTCAGGGTGGGCATCAAACTCAAACTACGCCCTGATCGGCGCACTGCGAGCAGTAGCCCAAACAATCTCATATGAAGTCACCCTAGCCATCATTCTACTATCAACATTACTAATGAGTGGCTCCTTTAACCTCTCCACCCTTATCACAACACAAGAACACCTCTGGTTACTCCTGCCATCATGGCCCTTGGCCATGATGTGGTTTATCTCCACACTAGCAGAGACCAACCGAACCCCCTTCGACCTTGCCGAAGGGGAGTCCGAACTAGTCTCAGGCTTCAACATCGAATACGCCGCAGGCCCCTTCGCCCTATTCTTCATGGCCGAATACACAAACATTATTATGATGAACACCCTCACCACTACAATCTTCCTAGGAACAACATATGACGCACTCTCCCCTGAACTCTACACAACATATTTTGTCACCAAGACCCTACTTCTAAACCTCCCTGTTCTTATGGATTCGAACAGCATACCCCCGATTCCGCTACGACCAACTCATGCACCTCCTATGGAAAAACTTCCTACCACTCACCCTAGCATTACTTATGTGGTATGTCTCCATGCCCATTACAATCTCCAGCATTCCCCCTCAAACCTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCAGCCCCTGTCCTCCCTTCACCCCCATTGCGTATGAGCATTTCAGAAACTCCAAGGAGTCACAGGCATCTTTATAGTTCACGTTAACATATAGACACTGTTGGAAGCAGTTCCTTCTAAAAGGGTAGCCCTGGACTTAATACCAGCCGGATACCTCTGGCCCCCACCCCATTACTGTACCTCTGGAGTCACTACTGTGGGTCGCCACTCCTCTGCTACACAGCACGGCTTTTTCAAGGCTGTATTGAGAAGGGAAGTTAGGAAGAAGGGTGTGCTGGGCTAACCAGCCCACAGAGCTCACATTCCTGTCCCTTGGGTGAAAAATACATGTCCATCCTGATATCTCCTGAAATTCAGAAAATTAGCCTCCACATGTGCAATGGCTTTAAGAGCCAGAAGCAGGGTTCTGGGAATTTTGCAAGTTACCTGTGGCCAGGTGTGGTCTCGGTTACCAAATACGGTTACCTGCAGCTTTTTAGTCCTTTGTGCTCCCACGGGTCTACAGAGTCCCATCTGCCCAAAGGTCTTGAAGCTTGACAGGATGTTTTCGATTACTCAGTCTCCCAGGGCACTACTGGTCCGTAGGATTCGATTGGTCGGGGTAGGAGAGTTAAACAACATTTAAACAGAGTTCTCTCAAAAATGTCTAAAGGGATTGTAGGTAGATAACATCCAATCACTGTTTGCACTTATCTGAAATCTTCCCTCTTGGCTGCCCCCAGGTATTTACTGTGGAGAACATTGCATAGGAATGTCTGGAAAAAGCTTCTACAACTTGTTACAGCCTTCACATTTGTAGAAGCTTT 40opt_COX10-ND1-3′UTR*ATGGCCGCCTCTCCACACACACTGAGTAGCAGACTGCTGACCGGCTGTGTTGGCGGCTCTGTGTGGTATCTGGAACGGCGGACAATGGCCAACCTCCTACTCCTCATTGTACCCATTCTAATCGCAATGGCATTCCTAATGCTTACCGAACGAAAAATTCTAGGCTATATGCAACTACGCAAAGGCCCCAACGTTGTAGGCCCCTACGGGCTACTACAACCCTTCGCTGACGCCATAAAACTCTTCACCAAAGAGCCCCTAAAACCCGCCACATCTACCATCACCCTCTACATCACCGCCCCGACCTTAGClCTCACCATCGCTCTTCTACTATGGACCCCCCTCCCCATGCCCAACCCCCTGGTCAACCTCAACCTAGGCCTCCTATTTATTCTAGCCACCTCTAGCCTAGCCGTTTACTCAATCCTCTGGTCAGGGTGGGCATCAAACTCAAACTACGCCCTGATCGGCGCACTGCGAGCAGTAGCCCAAACAATCTCATATGAAGTCACCCTAGCCATCATTCTACTATCAACATTACTAATGAGTGGCTCCTTTAACCTCTCCACCCTTATCACAACACAAGAACACCTCTGGTTACTCCTGCCATCATGGCCCTTGGCCATGATGTGGTTTATCTCCACACTAGCAGAGACCAACCGAACCCCCTTCGACCTTGCCGAAGGGGAGTCCGAACTAGTCTCAGGCTTCAACATCGAATACGCCGCAGGCCCCTTCGCCCTATTCTTCATGGCCGAATACACAAACATTATTATGATGAACACCCTCACCACTACAATCTTCCTAGGAACAACATATGACGCACTCTCCCCTGAACTCTACACAACATATTTTGTCACCAAGACCCTACTTCTAACCTCCCTGTTCTTATGGATTCGAACAGCATACCCCCGATTCCGCTACGACCAACTCATGCACCTCCTATGGAAAAACTTCCTACCACTCACCCTAGCATTACTTATGTGGTATGTCTCCATGCCCATTACAATCTCCAGCATTCCCCCTCAAACCTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGGTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCA 41 opt_COX10-opt_ND1-3′UTRATGGCCGCCTCTCCACACACACTGAGTAGCAGACTGCTGACCGGCTGTGTTGGCGGCTCTGTGTGGTATCTGGAACGGCGGACAATGGCCAACCTGCTGCTGCTGATCGTGCCCATCCTGATCGCCATGGCCTTCCTGATGCTGACCGAGCGCAAGATCCTGGGCTACATGCAGCTGCGCAAGGGCCCCAACGTGGTGGGCCCCTACGGCCTGCTGCAGCCCTTCGCCGACGCCATCAAGCTGTTCACCAAGGAGCCCCTGAAGCCCGCCACCAGCACCATCACCCTGTACATCACCGCCCCCACCCTGGCCCTGACCATCGCCCTGCTGCTGTGGACCCCCCTGCCCATGCCCAACCCCCTGGTGAACCTGAACCTGGGCCTGCTGTTCATCCTGGCCACCAGCAGCCTGGCCGTGTACAGCATCCTGTGGAGCGGCTGGGCCAGCAACAGCAACTACGCCCTGATCGGCGCCCTGCGCGCCGTGGCCCAGACCATCAGCTACGAGGTGACCCTGGCCATCATCCTGCTGAGCACCCTGCTGATGAGCGGCAGCTTCAACCTGAGCACCCTGATCACCACCCAGGAGCACCTGTGGCTGCTGCTGCCCAGCTGGCCCCTGGCCATGATGTGGTTCATCAGCACCCTGGCCGAGACCAACCGCACCCCCTTCGACCTGGCCGAGGGCGAGAGCGAGCTGGTGAGCGGCTTCAACATCGAGTACGCCGCCGGCCCCTTCGCCCTGTTCTTCATGGCCGAGTACACCAACATCATCATGATGAACACCCTGACCACCACCATCTTCCTGGGCACCACCTACGACGCCCTGAGCCCCGAGCTGTACACCACCTACTTCGTGACCAAGACCCTGCTGCTGACCAGCCTGTTCCTGTGGATCCGCACCGCCTACCCCCGCTTCCGCTACGACCAGCTGATGCACCTGCTGTGGAAGAACTTCCTGCCCCTGACCCTGGCCCTGCTGATGTGGTACGTGAGCATGCCCATCACCATCAGCAGCATCCCCCCCCAGACCTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCAGCCCCTGTCCTCCCTTCACCCCCATTGCGTATGAGCATTTCAGAACTCCAAGGAGTCACAGGCATCTTTATAGTTCACGTTAACATATAGACACTGTTGGAAGCAGTTCCTTCTAAAAGGGTAGCCCTGGACTTAATACCAGCCGGATACCTCTGGCCCCCACCCCATTACTGTACCTCTGGAGTCACTACTGTGGGTCGCCACTCCTCTGCTACACAGCACGGCTTTTTCAAGGCTGTATTGAGAAGGGAAGTTAGGAAGAAGGGTGTGCTGGGCTAACCAGCCCACAGAGCTCACATTCCTGTCCCTTGGGTGAAASATACATGTCCATCCTGATATCTCCTGAATTCAGAAATTAGCCTCCACATGTGCAATGGCTTTAAGAGCCAGAAGCAGGGTTCTGGGAATTTTGCAAGTTACCTGTGGCCAGGTGTGGTCTCGGTTACCAAATACGGTTACCTGCAGCTTTTTAGTCCTTTGTGCTCCCACGGGTCTACAGAGTCCCATCTGCCCAAAGGTCTTGAAGCTTGACAGGATGTTTTCGATTACTCAGTCTCCCAGGGCACTACTGGTCCGTAGGATTCGATTGGTCGGGGTAGGAGAGTTAAACAACATTTAAACAGAGTTCTCTCAAAAATGTCTAAAGGGATTGTAGGTAGATAACATCCAATCACTGTTTGCACTTATCTGAAATCTTCCCTCTTGGCTGCCCCCAGGTATTTACTGTGGAGAACATTGCATAGGAATGTCTGGAAAAAGCTTCTACAACTTGTTACAGCCTTCACATTTGTAGAAGCTTT 42 opt_COX10-opt_ND1-3′UTRATGGCCGCCTCTCCACACACACTGAGTAGCAGACTGCTGACCGGCTGTGTTGGCGGCTCTGTGTGGTATCTGGAACGGCGGACAATGGCCAACCTGCTGCTGCTGATCGTGCCCATCCTGATCGCCATGGCCTTCCTGATGCTGACCGAGCGCAAGATCCTGGGCTACATGCAGCTGCGCAAGGGCCCCAACGTGGTGGGCCCCTACGGCCTGCTGCAGCCCTTCGCCGACGCCATCAAGCTGTTCACCAAGGAGCCCCTGAAGCCCGCCACCAGCACCATCACCCTGTACATCACCGCCCCCACCCTGGCCCTGACCATCGCCCTGCTGCTGTGGACCCCCCTGCCCATGCCCAACCCCCTGGTGAACCTGAACCTGGGCCTGCTGTTCATCCTGGCCACCAGCAGCCTGGCCGTGTACAGCATCCTGTGGAGCGGCTGGGCCAGCAACAGCAACTACGCCCTGATCGGCGCCCTGCGCGCCGTGGCCCAGACCATCAGCTACGAGGTGACCCTGGCCATCATCCTGCTGAGCACCCTGCTGATGAGCGGCAGCTTCAACCTGAGCACCCTGATCACCACCCAGGAGCACCTGTGGCTGCTGCTGCCCAGCTGGCCCCTGGCCATGATGTGGTTCATCAGCACCCTGGCCGAGACCAACCGCACCCCCTTCGACCTGGCCGAGGGCGAGAGCGAGCTGGTGAGCGGCTTCAACATCGAGTACGCCGCCGGCCCCTTCGCCCTGTTCTTCATGGCCGAGTACACCAACATCATCATGATGAACACCCTGACCACCACCATCTTCCTGGGCACCACCTACGACGCCCTGAGCCCCGAGCTGTACACCACCTACTTCGTGACCAAGACCCTGCTGCTGACCAGCCTGTTCCTGTGGATCCGCACCGCCTACCCCCGCTTCCGCTACGACCAGCTGATGCACCTGCTGTGGAAGAACTTCCTGCCCCTGACCCTGGCCCTGCTGATGTGGTACGTGAGCATGCCCATCACCATCAGCAGCATCCCCCCCCAGACCTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCA 43 opt_COX10*-ND4-3′UTRATGGCCGCCAGCCCCCACACCCTGAGCAGCCGCCTGCTGACCGGCTGCGTGGGCGGCAGCGTGTGGTACCTGGAGCGCCGCACCATGCTAAAACTAATCGTCCCAACAATTATGTTACTACCACTGACATGGCTTTCCAAAAAACACATGATTTGGATCAACACAACCACCCACAGCCTAATTATTAGCATCATCCCTCTACTATTTTTTAACCAAATCAACAACAACCTATTTAGCTGTTCCCCAACCTTTTCCTCCGACCCCCTAACAACCCCCCTCCTAATGCTAACTACCTGGCTCCTACCCCTCACAATCATGGCAAGCCAACGCCACTTATCCAGTGAACCACTATCACGAAAAAAACTCTACCTCTCTATGCTAATCTCCCTACAAATCTCCTTAATTATGACATTCACAGCCACAAACTAATCATGTTTTATATCTTCTTCGAAACCACACTTATCCCCACCTTGGCTATCATCACCCGATGGGGCAACCAGCCAGAACGCCTGAACGCAGGCACATACTTCCTATTCTACACCCTAGTAGGCTCCCTTCCCCTACTCATCGCACTAATTTACACTCACAACACCCTAGGCTCACTAAACATTCTACTACTCACTCTCACTGCCCAAGAACTATCAAACTCCTGGGCCAACAACTTAATGTGGCTAGCTTACACAATGGCTTTTATGGTAAAGATGCCTCTTTACGGACTCCACTTATGGCTCCCTAAAGCCCATGTCGAAGCCCCCATCGCTGGGTCAATGGTACTTGCCGCAGTACTCTTAAAACTAGGCGGCTATGGTATGATGCGCCTCACACTCATTCTCAACCCCCTGACAAAACACATGGCCTACCCCTTCCTTGTACTATCCCTATGGGGCATGATTATGACAAGCTCCATCTGCCTACGACAAACAGACCTAAAATCGCTCATTGCATACTCTTCAATCAGCCACATGGCCCTCGTAGTAACAGCCATTCTCATCCAAACCCCCTGGAGCTTCACCGGCGCAGTCATTCTCATGATCGCCCACGGGCTTACATCCTCATTACTATTCTGCCTAGCAAACTCAAACTACGAACGCACTCACAGTCGCATCATGATCCTCTCTCAAGGACTTCAAACTCTACTCCCACTAATGGCTTTTTGGTGGCTTCTAGCAAGCCTCGCTAACCTCGCCTTACCCCCCACTATTAACCTACTGGGAGAACTCTCTGTGCTAGTAACCACGTTCTCCTGGTCAAATATCACTCTCCTACTTACAGGACTCAACATGCTAGTCACAGCCCTATACTCCCTCTACATGTTTACCACAACACAATGGGGCTCACTCACCCACCACATTAACAACATGAAACCCTCATTCACACGAGAAAACACCCTCATGTTCATGCACCTATCCCCCATTCTCCTCCTATCCCTCAACCCCGACATCATTACCGGGTTTTCCTCTTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCAGCCCCTGTCCTCCCTTCACCCCCATTGCGTATGAGCATTTCAGAACTCCAAGGAGTCACAGGCATCTTTATAGTTCACGTTAACATATAGACACTGTTGGAAGCAGTTCCTTCTAAAAGGGTAGCCCTGGACTTAATACCAGCCGGATACCTCTGGCCCCCACCCCATTACTGTACCTCTGGAGTCACTACTGTGGGTCGCCACTCCTCTGCTACACAGCACGGCTTTTTCAAGGCTGTATTGAGAAGGGAAGTTAGGAAGAAGGGTGTGCTGGGCTAACCAGCCCACAGAGCTCACATTCCTGTCCCTTGGGTGAAAAATACATGTCCATCCTGATATCTCCTGAATTCAGAAATTAGCCTCCACATGTGCAATGGCTTTAAGAGCCAGAAGCAGGGTTCTGGGAATTTTGCAAGTTACCTGTGGCCAGGTGTGGTCTCGGTTACCAAATACGGTTACCTGCAGCTTTTTAGTCCTTTGTGCTCCCACGGGTCTACAGAGTCCCATCTGCCCAAAGGTCTTGAAGCTTGACAGGATGTTTTCGATTACTCAGTCTCCCAGGGCACTACTGGTCCGTAGGATTCGATTGGTCGGGGTAGGAGAGTTAAACAACATTTAAACAGAGTTCTCTCAAAAATGTCTAAAGGGATTGTAGGTAGATAACATCCAATCACTGTTTGCACTTATCTGAAATCTTCCCTCTTGGCTGCCCCCAGGTATTTACTGTGGAGAACATTGCATAGGAATGTCTGGAAAAAGCTTCTACAACTTGTTACAGCCTTCACATTTGTAGAAGCTTT 44 opt_COX10*-ND4-3′UTR*ATGGCCGCCAGCCCCCACACCCTGAGCAGCCGCCTGCTGACCGGCTGCGTGGGCGGCAGCGTGTGGTACCTGGAGCGCCGCACCATGCTAAAACTAATCGTCCCAACAATTATGTTACTACCACTGACATGGCTTTCCAAAAAACACATGATTTGGATCAACACAACCACCCACAGCCTAATTATTAGCATCATCCCTCTACTATTTTTTAACCAAATCAACAACAACCTATTTAGCTGTTCCCCAACCTTTTCCTCCGACCCCCTAACAACCCCCCTCCTAATGCTAACTACCTGGCTCCTACCCCTCACAATCATGGCAAGCCAACGCCACTTATCCAGTGAACCACTATCACGAAAAAAACTCTACCTCTCTATGCTAATCTCCCTACAAATCTCCTTAATTATGACATTCACAGCCACAGAACTAATCATGTTTTATATCTTCTTCGAAACCACACTTATCCCCACCTTGGCTATCATCACCCGATGGGGCAACCAGCCAGAACGCCTGAACGCAGGCACATACTTCCTATTCTACACCCTAGTAGGCTCCCTTCCCCTACTCATCGCACTAATTTACACTCACAACACCCTAGGCTCACTAAACATTCTACTACTCACTCTCACTGCCCAAGAACTATCAAACTCCTGGGCCAACAACTTAATGTGGCTAGCTTACACAATGGCTTTTATGGTAAAGATGCCTCTTTACGGACTCCACTTATGGCTCCCTAAAGCCCATGTCGAAGCCCCCATCGCTGGGTCAATGGTACTTGCCGCAGTACTCTTAAAACTAGGCGGCTATGGTATGATGCGCCTCACACTCATTCTCAACCCCCTGACAAAACACATGGCCTACCCCTTCCTTGTACTATCCCTATGGGGCATGATTATGACAAGCTCCATCTGCCTACGACAAACAGACCTAAAATCGCTCATTGCATACTCTTCAATCAGCCACATGGCCCTCGTAGTAACAGCCATTCTCATCCAAACCCCCTGGAGCTTCACCGGCGCAGTCATTCTCATGATCGCCCACGGGCTTACATCCTCATTACTATTCTGCCTAGCAAACTCAAACTACGAACGCACTCACAGTCGCATCATGATCCTCTCTCAAGGACTTCAAACTCTACTCCCACTAATGGCTTTTTGGTGGCTTCTAGCAAGCCTCGCTAACCTCGCCTTACCCCCCACTATTbACCTACTGGGAGAACTCTCTGTGCTAGTAACCACGTTCTCCTGGTCAAATATCACTCTCCTACTTACAGGACTCAACATGCTAGTCACAGCCCTATACTCCCTCTACATGTTTACCACAACACAATGGGGCTCACTCACCCACCACATTAACAACATGAAACCCTCATTCACACGAGAAAACACCCTCATGTTCATGCACCTATCCCCCATTCTCCTCCTATCCCTCAACCCCGACATCATTACCGGGTTTTCCTCTTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCA 45 opt_COX10*-opt_ND4-3′UTRATGGCCGCCAGCCCCCACACCCTGAGCAGCCGCCTGCTGACCGGCTGCGTGGGCGGCAGCGTGTGGTACCTGGAGCGCCGCACCATGCTGAAGCTGATCGTGCCCACCATCATGCTGCTGCCTCTGACCTGGCTGAGCAAGAAACACATGATCTGGATCAACACCACCACGCACAGCCTGATCATCAGCATCATCCCTCTGCTGTTCTTCAACCAGATCAACAACAACCTGTTCAGCTGCAGCCCCACCTTCAGCAGCGACCCTCTGACAACACCTCTGCTGATGCTGACCACCTGGCTGCTGCCCGTCACAATCATGGCCTCTCAGAGACACCTGAGCAGCGAGCCCGTGAGCCGGAAGAAACTGTACCTGAGCATGCTGATCTCCCTGCAGATCTCTCTGATCATGACCTTCACCGCCACCGAGCTGATCATGTTCTACATCTTTTTCGAGACAACGCTGATCCCCACACTGGCCATCATCACCAGATGGGGCAACCAGCCTGAGAGACTGAACGCCGGCACCTACTTTCTGTTCTACACCCTCGTGGGCAGCCTGCCACTGCTGATTGCCCTGATCTACACCCACAACACCCTGGGCTCCCTGAACATCCTGCTGCTGACACTGACAGCCCAAGAGCTGAGCAACAGGTGGGCCAACAATCTGATGTGGCTGGCCTACACAATGGCCTTCATGGTCAAGATGCCCCTGTACGGCCTGCACCTGTGGCTGCCTAAAGCTCATGTGGAAGCCCCTATCGCCGGCTCTATGGTGCTGGCTGCAGTGCTGCTGAACTCGGCGGCTACGGCATGATGCGGCTGACCCTGATTCTGAATCCCCTGACCAAGCACATGGCCTATCCATTTCTGGTGCTGAGCCTGTGGGGCATGATTATGACCAGCAGCATCTGCCTGCGGCAGACCGATCTGAAGTCCCTGATCGCCTACAGCTCCATCAGCCACATGGCCCTGGTGGTCACCGCCATCCTGATTCAGACCCCTTGGAGCTTTACAGGCGCCGTGATCCTGATGATTGCCCACGGCCTGACAAGCAGCCTGCTGTTTTGTCTGGCCAACAGCAACTACGAGCGGACCCACAGCAGAATCATGATCCTGTCTCAGGGCCTGCAGACCCTCCTGCCTCTTATGGCTTTTTGGTGGCTGCTGGCCTCTCTGGCCAATCTGGCACTGCCTCCTACCATCAATCTGCTGGGCGAGCTGAGCGTGCTGGTCACCACATTCAGCTGGTCCAATTATCACCCTGCTGCTCACCGGCCTGAACATGCTGGTTACAGCCCTGTACTCCCTGTACATGTTCACCACCACACAGTGGGGAAGCCTGACACACCACATCAACAATATGAAGCCCAGCTTCACCCGCGAGAACACCCTGATGTTCATGCATCTGAGCCCCATTCTGCTGCTGTCCCTGAATCCTGATATCATCACCGGCTTCTCCAGCTGAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCAGCCCCTGTCCTCCCTTCACCCCCATTGCGTATGAGCATTTCAGAACTCCAAGGAGTCACAGGCATCTTTATAGTTCACGTTAACATATAGACACTGTTGGAAGCAGTTCCTTCTAAAAGGGTAGCCCTGGACTTAATACCAGCCGGATACCTCTGGCCCCCACCCCATTACTGTACCTCTGGAGTCACTACTGTGGGTCGCCACTCCTCTGCTACACAGCACGGCTTTTTCAAGGCTGTATTGAGAAGGGAAGTTAGGAAGAAGGGTGTGCTGGGCTAACCAGCCCACAGAGCTCACATTCCTGTCCCTTGGGTGAAAAATACATGTCCATCCTGATATCTCCTGAATTCAGAAATTAGCCTCCACATGTGCAATGGCTTTAAGAGCCAGAAGCAGGGTTCTGGGAATTTTGCAAGTTACCTGTGGCCAGGTGTGGTCTCGGTTACCAAATACGGTTACCTGCAGCTTTTTAGTCCTTTGTGCTCCCACGGGTCTACAGAGTCCCATCTGCCCAAAGGTCTTGAAGCTTGACAGGATGTTTTCGATTACTCAGTCTCCCAGGGCACTACTGGTCCGTAGGATTCGATTGGTCGGGGTAGGAGAGTTAAACAACATTTAAACAGAGTTCTCTCAAAAATGTCTAAAGGGATTGTAGGTAGATAACATCCAATCACTGTTTGCACTTATCTGAAATCTTCCCTCTTGGCTGCCCCCAGGTATTTACTGTGGAGAACATTGCATAGGAATGTCTGGAAAAAGCTTCTACAACTTGTTACAGCCTTCACATTTGTAGAAGCTTT 46 opt_COX10*-opt_ND4-3′UTR*ATGGCCGCCAGCCCCCACACCCTGAGCAGCCGCCTGCTGACCGGCTGCGTGGGCGGCAGCGTGTGGTACCTGGAGCGCCGCACCATGCTGAAGCTGATCGTGCCCACCATCATGCTGCTGCCTCTGACCTGGCTGAGCAAGAAACACATGATCTGGATCAACACCACCACGCACAGCCTGATCATCAGCATCATCCCTCTGCTGTTCTTCAACCAGATCAACAACAACCTGTTCAGCTGCAGCCCCACCTTCAGCAGCGACCCTCTGACAACACCTCTGCTGATGCTGACCACCTGGCTGCTGCCCCTCACAATCATGGCCTCTCAGAGACACCTGAGCAGCGAGCCCCTGAGCCGGAAGAAACTGTACCTGAGCATGCTGATCTCCCTGCAGATCTCTCTGATCATGACCTTCACCGCCACCGAGCTGATCATGTTCTACATCTTTTTCGAGACAACGCTGATCCCCACACTGGCCATCATCACCAGATGGGGCAACCAGCCTGAGAGACTGAACGCCGGCACCTACTTTCTGTTCTACACCCTCGTGGGCAGCCTGCCACTGCTGATTGCCCTGATCTACACCCACAACACCCTGGGCTCCCTGAACATCCTGCTGCTGACACTGACAGCCCAAGAGCTGAGCAACAGCTGGGCCAACAATCTGATGTGGCTGGCCTACACAATGGCCTTCATGGTCAAGATGCCCCTGTACGGCCTGCACCTGTGGCTGCCTAAAGCTCATGTGGAAGCCCCTATCGCCGGCTCTATGGTGCTGGCTGCAGTGCTGCTGAAACTCGGCGGCTACGGCATGATGCGGCTGACCCTGATTCTGAATCCCCTGACCAAGCACATGGCCTATCCATTTCTGGTGCTGAGCCTGTGGGGCATGATTATGACCAGCAGCATCTGCCTGCGGCAGACCGATCTGAAGTCCCTGATCGCCTACAGCTCCATCAGCCACATGGCCCTGGTGGTCACCGCCATCCTGATTCAGACCCCTTGGAGCTTTACAGGCGCCGTGATCCTGATGATTGCCCACGGCCTGACAAGCAGCCTGCTGTTTTGTCTGGCCAACAGCAACTACGAGCGGACCCACAGCAGAATCATGATCCTGTCTCAGGGCCTGCAGACCCTCCTGCCTCTTATGGCTTTTTGGTGGCTGCTGGCCTCTCTGGCCAATCTGGCACTGCCTCCTACCATCAATCTGCTGGGCGAGCTGAGCGTGCTGGTCACCACATTCAGCTGGTCCAATATCACCCTGCTGCTCACCGGCCTGAACATGCTGGTTACAGCCCTGTACTCCCTGTACATGTTCACCACCACACAGTGGGGAAGCCTGACACACCACATCAACAATATGAAGCCCAGCTTCACCCGCGAGAACACCCTGATGTTCATGCATCTGAGCCCCATTCTGCTGCTGTCCCTGAATCCTGATATCATCACCGGCTTCTCCAGCTGAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCA 47opt_COX10*-opt_ND4*-3′UTRATGGCCGCCAGCCCCCACACCCTGAGCAGCCGCCTGCTGACCGGCTGCGTGGGCGGCAGCGTGTGGTACCTGGAGCGCCGCACCATGCTGAAGCTGATCGTGCCCACCATCATGCTGCTGCCCCTGACCTGGCTGAGCAAGAAGCACATGATCTGGATCAACACCACCACCCACAGCCTGATCATCAGCATCATCCCCCTGCTGTTCTTCAACCAGATCAACAACAACCTGTTCAGCTGCAGCCCCACCTTCAGCAGCGACCCCCTGACCACCCCCCTGCTGATGCTGACCACCTGGCTGCTGCCCCTGACCATCATGGCCAGCCAGCGCCACCTGAGCAGCGAGCCCCTGAGCCGCAAGAAGCTGTACCTGAGCATGCTGATCAGCCTGCAGATCAGCCTGATCATGACCTTCACCGCCACCGAGCTGATCATGTTCTACATCTTCTTCGAGACCACCCTGATCCCCACCCTGGCCATCATCACCCGCTGGGGCAACCAGCCCGAGCGCCTGAACGCCGGCACCTACTTCCTGTTCTACACCCTGGTGGGCAGCCTGCCCCTGCTGATCGCCCTGATCTACACCCACAACACCCTGGGCAGCCTGAACATCCTGCTGCTGACCCTGACCGCCCAGGAGCTGAGCAACAGCTGGGCCAACAACCTGATGTGGCTGGCCTACACCATGGCCTTCATGGTGAAGATGCCCCTGTACGGCCTGCACCTGTGGCTGCCCAAGGCCCACGTGGAGGCCCCCATCGCCGGCAGCATGGTGCTGGCCGCCGTGCTGCTGAAGCTGGGCGGCTACGGCATGATGCGCCTGACCCTGATCCTGAACCCCCTGACCAAGCACATGGCCTACCCCTTCCTGGTGCTGAGCCTGTGGGGCATGATCATGACCAGCAGCATCTGCCTGCGCCAGACCGACCTGAAGAGCCTGATCGCCTACAGCAGCATCAGCCACATGGCCCTGGTGGTGACCGCCATCCTGATCCAGACCCCCTGGAGCTTCACCGGCGCCGTGATCCTGATGATCGCCCACGGCCTGACCAGCAGCCTGCTGTTCTGCCTGGCCAACAGCAACTACGAGCGCACCCACAGCCGCATCATGATCCTGAGCCAGGGCCTGCAGACCCTGCTGCCCCTGATGGCCTTCTGGTGGCTGCTGGCCAGCCTGGCCAACCTGGCCCTGCCCCCCACCATCAACCTGCTGGGCGAGCTGAGCGTGCTGGTGACCACCTTCAGCTGGAGCAACATCACCCTGCTGCTGACCGGCCTGAACATGCTGGTGACCGCCCTGTACAGCCTGTACATGTTCACCACCACCCAGTGGGGCAGCCTGACCCACCACATCAACAACATGAAGCCCAGCTTCACCCGCGAGAACACCCTGATGTTCATGCACCTGAGCCCCATCCTGGTGCTGAGCCTGAACCCCGACATCATCACCGGCTTCAGCAGCTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCAGCCCCTGTCCTCCCTTCACCCCCATTGCGTATGAGCATTTCAGAACTCCAAGGAGTCACAGGCATCTTTATAGTTCACGTTAACATATAGACACTGTTGGAAGCAGTTCCTTCTAAAAGGGTAGCCCTGGACTTAATACCAGCCGGATACCTCTGGCCCCCACCCCATTACTGTACCTCTGGAGTCACTACTGTGGGTCGCCACTCCTCTGCTACACAGCACGGCTTTTTCAAGGCTGTATTGAGAAGGGAAGTTAGGAAGAAGGGTGTGCTGGGCTbACCAGCCCACAGAGCTCACATTCCTGTCCCTTGGGTGAAAAATACATGTCCATCCTGATATCTCCTGAATTCAGAAATTAGCCTCCACATGTGCAATGGCTTTAAGAGCCAGAAGCAGGGTTCTGGGAATTTTGCAAGTTACCTGTGGCCAGGTGTGGTCTCGGTTACCAAATACGGTTACCTGCAGCTTTTTAGTCCTTTGTGCTCCCACGGGTCTACAGAGTCCCAlCTGCCCAAAGGTCTTGAAGCTTGACAGGATGTTTTCGATTACTCAGTCTCCCAGGGCACTACTGGTCCGTAGGATTCGATTGGTCGGGGTAGGAGAGTTAAACAACATTTAAACAGAGTTCTCTCAAAAATGTCTAAAGGGATTGTAGGTAGATAACATCCAATCACTGTTTGCACTTATCTGAAATCTTCCCTCTTGGCTGCCCCCAGGTATTTACTGTGGAGAACATTGCATAGGAATGTCTGGAAAAAGCTTCTACAACTTGTTACAGCCTTCACATTTGTAGAAGCTTT 48opt_COX10*-opt_ND4*-3′UTRATGGCCGCCAGCCCCCACACCCTGAGCAGCCGCCTGCTGACCGGCTGCGTGGGCGGCAGCGTGTGGTACCTGGAGCGCCGCACCATGCTGAAGCTGATCGTGCCCACCATCATGCTGCTGCCCCTGACCTGGCTGAGCAAGAAGCACATGATCTGGATCAACACCACCACCCACAGCCTGATCATCAGCATCATCCCCCTGCTGTTCTTCAACCAGATCAACAACAACCTGTTCAGCTGCAGCCCCACCTTCAGCAGCGACCCCCTGACCACCGCCCTGCTGATGCTGACCACCTGGCTGCTGCCCCTGACCATCATGGCCAGCCAGCGCCACCTGAGCAGCGAGCCCCTGAGCCGCAAGAAGCTGTACCTGAGCATGCTGATCAGCCTGCAGATCAGCCTGATCATGACCTTCACCGCCACCGAGCTGATCATGTTCTACATCTTCTTCGAGACCACCCTGATCCCCACCCTGGCCATCATCACCCGCTGGGGCAACCAGCCCGAGCGCCTGAACGCCGGCACCTACTTCCTGTTCTACACCCTGGTGGGCAGCCTGCCCCTGCTGATCGCCCTGATCTACACCCACAACACCCTGGGCAGCCTGAACATCCTGCTGCTGACCCTGACCGCCCAGGAGCTGAGCAACAGCTGGGCCAACAACCTGATGTGGCTGGCCTACACCATGGCCTTCATGGTGAAGATGCCCCTGTACGGCCTGCACCTGTGGCTGCCCAAGGCCCACGTGGAGGCCCCCATCGCCGGCAGCATGGTGCTGGCCGCCGTGCTGCTGAAGCTGGGCGGCTACGGCATGATGCGCCTGACCCTGATCCTGAACCGCCTGACCAAGCACATGGCCTACCCGTTCCTGGTGCTGAGCCTGTGGGGCATGATCATGACCAGCAGCATCTGCCTGCGCCAGACCGACCTGAAGAGCCTGATCGCCTACAGCAGCATCAGCCACATGGCCCTGGTGGTGACCGCCATCCTGATCCAGACCCCCTGGAGCTTCACCGGCGCCGTGATCCTGATGATCGCCCACGGCCTGACCAGCAGCCTGCTGTTCTGCCTGGCCAACAGCAACTACGAGCGCACCCACAGCCGCATCATGATCCTGAGCCAGGGCCTGGAGACCCTGCTGCCCCTGATGGCCTTCTGGTGGCTGCTGGCCAGCCTGGCCAACCTGGCCCTGCCCCCCACCATCAACCTGCTGGGCGAGCTGAGCGTGCTGGTGACCACCTTCAGCTGGAGCAACATCACCCTGCTGCTGACCGGCCTGAACATGCTGGTGACCGCCCTGTACAGCCTGTACATGTTCACCACCACCCAGTGGGGCAGCCTGACCCACCACATCAACAACATGAAGCCCAGCTTCACCCGCGAGAACACCCTGATGTTCATGCACCTGAGCCCCATCCTGGTGCTGAGCCTGAACCCCGACATCATCACCGGCTTCAGCAGCTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCCATAATGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTG CCA49 opt_COX10*-ND6-3′UTRATGGCCGCCAGCCCCCACACCCTGAGCAGCCGCCTGCTGACCGGCTGCGTGGGCGGCAGCGTGTGGTACCTGGAGCGCCGCACCATGATGTATGCTTTGTTTCTGTTGAGTGTGGGTTTAGTAATGGGGTTTGTGGGGTTTTCTTCTAAGCCTTCTCCTATTTATGGGGGTTTAGTATTGATTGTTAGCGGTGTGGTCGGGTGTGTTATTATTCTGAATTTTGGGGGAGGTTATATGGGTTTAATGGTTTTTTTAATTTATTTAGGGGGAATGATGGTTGTCTTTGGATATACTACAGCGATGGGTATTGAGGAGTATCCTGAGGCATGGGGGTCAGGGGTTGAGGTCTTGGTGAGTGTTTTAGTGGGGTTAGCGATGGAGGTAGGATTGGTGCTGTGGGTGAAAGAGTATGATGGGGTGGTGGTTGTGGTAAACTTTAATAGTGTAGGAAGCTGGATGATTTATGAAGGAGAGGGGTCAGGGTTGATTCGGGAGGATCCTATTGGTGCGGGGGCTTTGTATGATTATGGGCGTTGGTTAGTAGTAGTTACTGGTTGGACATTGTTTGTTGGTGTATATATTGTAATTGAGATTGGTCGGGGGAATTAGGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCAGCCCCTGTCCTCCCTTCACCCCCATTGCGTATGAGCATTTCAGAACTCCAAGGAGTCACAGGCATCTTTATAGTTCACGTTAACATATAGACACTGTTGGAAGCAGTTCCTTCTAAAAGGGTAGCCCTGGACTTAATACCAGCCGGATACCTCTGGCCCCCACCCCATTACTGTACCTCTGGAGTCACTACTGTGGGTCGCCACTCCTCTGCTACACAGCACGGCTTTTTCAAGGCTGTATTGAGAAGGGAAGTTAGGAAGAAGGGTGTGCTGGGCTAACCAGCCCACAGAGCTCACATTCCTGTCCCTTGGGTGAAAAATACATGTCCATCCTGATATCTCCTGAATTCAGAAATTAGCCTCCACATGTGCAATGGCTTTAAGAGCCAGAAGCAGGGTTCTGGGAATTTTGCAAGTTACCTGTGGCCAGGTGTGGTCTCGGTTACCAAATACGGTTACCTGCAGCTTTTTAGTCCTTTGTGCTCCCACGGGTCTACAGAGTCCCATCTGCCCAAAGGTCTTGAAGGTTGACAGGATGTTTTCGATTACTCAGTCTCCCAGGGCACTACTGGTCCGTAGGATTCGATTGGTCGGGGTAGGAGAGTTAAACAACATTTAAACAGAGTTCTCTCAAAAATGTCTAAAGGGATTGTAGGTAGATAACATCCAATCACTGTTTGCACTTATCTGAAATCTTCCCTCTTGGCTGCCCCCAGGTATTTACTGTGGAGAACATTGCATAGGAATGTCTGGAAAAAGCTTCTACAACTTGTTACAGCCTTCACATTTGTAGAAGGTTT 50opt_COX10*-ND6-3′UTR*ATGGCCGCCAGCCCCCACACCCTGAGCAGCCGCCTGCTGACCGGCTGCGTGGGCGGCAGCGTGTGGTACCTGGAGCGCCGCACCATGATGTATGCTTTGTTTCTGTTGAGTGTGGGTTTAGTAATGGGGTTTGTGGGGTTTTCTTCTAAGCCTTCTCCTATTTATGGGGGTTTAGTATTGATTGTTAGCGGTGTGGTCGGGTGTGTTATTATTCTGAATTTTGGGGGAGGTTATATGGGTTTAATGGTTTTTTTAATTTATTTAGGGGGAATGATGGTTGTCTTTGGATATACTACAGCGATGGCTATTGAGGAGTATCCTGAGGCATGGGGGTCAGGGGTTGAGGTCTTGGTGAGTGTTTTAGTGGGGTTAGCGATGGAGGTAGGATTGGTGGTGTGGGTGAAAGAGTATGATGGGGTGGTGGTTGTGGTAAACTTTAATAGTGTAGGAAGCTGGATGATTTATGAAGGAGAGGGGTCAGGGTTGATTCGGGAGGATCCTATTGGTGCGGGGGCTTTGTATGATTATGGGCGTTGGTTAGTAGTAGTTACTGGTTGGACATTGTTTGTTGGTGTATATATTGTAATTGAGATTGCTCGGGGGAATTAGGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCA 51 opt_COX10*-opt_ND6-3′UTRATGGCCGCCAGCCCCCACACCCTGAGCAGCCGCCTGCTGACCGGCTGCGTGGGCGGCAGCGTGTGGTACCTGGAGCGCCGCACCATGATGTACGCCCTGTTCCTGCTGAGCGTGGGCCTGGTGATGGGCTTCGTGGGCTTCAGCAGCAAGCCCAGCCCCATCTACGGCGGCCTGGTGCTGATCGTGAGCGGCGTGGTGGGCTGCGTGATCATCCTGAACTTCGGCGGCGGCTACATGGGCCTGATGGTGTTCCTGATCTACCTGGGCGGCATGATGGTGGTGTTCGGCTACACCACCGCCATGGCCATCGAGGAGTACCCCGAGGCCTGGGGCAGCGGCGTGGAGGTGCTGGTGAGCGTGCTGGTGGGCCTGGCCATGGAGGTGGGCCTGGTGCTGTGGGTGAAGGAGTACGACGGCGTGGTGGTGGTGGTGAACTTCAACAGCGTGGGCAGCTGGATGATCFACGAGGGCGAGGGCAGCGGCCTGATCCGCGAGGACCCCATCGGCGCCGGCGCCCTGTACGACTACGGCCGCTGGCTGGTGGTGGTGACCGGCTGGACCCTGTTCGTGGGCGTGTACATCGTGATCGAGATCGCCCGCGGCAACTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCAGCCCCTGTCCTCCCTTCACCCCCATTGCGTATGAGCATTTCAGAACTCCAAGGAGTCACAGGCATCTTTATAGTTCACGTTAACATATAGACACTGTTGGAAGCAGTTCCTTCTAAAAGGGTAGCCCTGGACTTAATACCAGCCGGATACCTCTGGCCCCCACCCCATTACTGTACCTCTGGAGTCACTACTGTGGGTCGCCACTCCTCTGCTACACAGCACGGCTTTTTCAAGGCTGTATTGAGAAGGGAAGTTAGGAAGAAGGGTGTGCTGGGCTAACCAGCCCACAGAGCTCACATTCCTGTCCCTTGGGTGAAAAATACATGTCCATCCTGATATCTCCTGAATTCAGAAATTAGCCTCCACATGTGCAATGGCTTTAAGAGCCAGAAGCAGGGTTTCTGGGAATTTTGCAAGTTACCTGTGGCCAGGTGTGGTCTCGGTTACCAAATACGGTTACCTGCAGCTTTTTAGTCCTTTGTGCTCCCACGGGTCTACAGAGTCCCATCTGCCCAASAGGTCTTGAAGCTTGACAGGATGTTTTCGATTACTCAGTCTCCCAGGGCACTACTGGTCCGTAGGATTCGATTGGTCGGGGTAGGAGAGTTAAACAACATTTAAACAGAGTTCTCTCAAAAATGTCTAAAGGGATTGTAGGTAGATAACATCCAATCACTGTTTGCACTTATCTGAAATCCTCTTGGCTGCCCCCAGGTATTTACTGTGGAGAACATTGCATAGGAATGTCTGGAAAAAGCTTCTACAACTTGTTACAGCCTTCACATTTGTAGAAAGCTTT 52 opt_COX10*-opt_ND6-3′UTR*ATGGCCGCCAGCCCCCACACCCTGAGCAGCCGCCTGCTGACCGGCTGCGTGGGCGGCAGCGTGTGGTACCTGGAGCGCCGCACCATGATGTACGCCCTGTTCCTGCTGAGCGTGGGCCTGGTGATGGGCTTCGTGGGCTTCAGCAGCAAGCCCAGCCCCATCTACGGCGGCCTGGTGCTGATCGTGAGCGGCGTGGTGGGCTGCGTGATCATCCTGAACTTCGGCGGCGGCTACATGGGCCTGATGGTGTTCCTGATCTACCTGGGCGGCATGATGGTGGTGTTCGGCTACACCACCGCCATGGCCATCGAGGAGTACCCCGAGGCCTGGGGCAGCGGCGTGGAGGTGCTGGTGAGCGTGCTGGTGGGCCTGGCCATGGAGGTGGGCCTGGTGCTGTGGGTGAAGGAGTACGACGGCGTGGTGGTGGTGGTGAACTTCAACAGCGTGGGCAGCTGGATGATCTACGAGGGCGAGGGCAGCGGCCTGATCCGCGAGGACCCCATCGGCGCCGGCGCCCTGTACGACTACGGCCGCTGGCTGGTGGTGGTGACCGGCTGGACCCTGTTCGTGGGCGTGTACATCGTGATCGAGATCGCCCGCGGCAACTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCA 53 opt_COX10*-opt_ND6-3′UTR*ATGGCCGCCAGCCCCCACACCCTGAGCAGCCGCCTGCTGACCGGCTGCGTGGGCGGCAGCGTGTGGTACCTGGAGCGCCGCACCATGGCCAACCTCCTACTCCTCATTGTACCCATTCTAATCGCAATGGCATTCCTAATGCTTACCGAACGAAAAATTCTAGGCTATATGCAACTACGCAAAGGCCCCAACGTTGTAGGCCCCTACGGGCTACTACAACCCTTCGCTGACGCCATAAAACTCTTCACCAAAGAGCCCCTAAAACCCGCCACATCTACCATCACCCTCTACATCACCGCCCCGACCTTAGCTCTCACCATCGCTCTTCTACTATGGACCCCCCTCCCCATGCCCAACCCCCTGGTCAACCTCAACCTAGGCCTCCTATTTATTCTAGCCACCTCTAGCCTAGCCGTTTACTCAATCCTCTGGTCAGGGTGGGCATCAAACTCAAACTACGCCCTGATCGGCGCACTGCGAGCAGTAGCCCAAACAATCTCATATGAAGTCACCCTAGCCATCATTCTACTATCAACATTACTAATGAGTGGCTCCTTTAACCTCTCCACCCTTATCACAACACAAGAACACCTCTGGTTACTCCTGCCATCATGGCCCTTGGCCATGATGTGGTTTATCTCCACACTAGCAGAGACCAACCGAACCCCCTTCGACCTTGCCGAAGGGGAGTCCGAACTAGTCTCAGGCTTCAACATCGAATACGCCGCAGGCCCCTTCGCCCTATTCTTCATGGCCGAATACACAAACATTATTATGATGAACACCCTCACCACTACAATCTTCCTAGGAACAACATATGACGCACTCTCCCCTGAACTCTACACAACATATTTTGTCACCAAGACCCTACTTCTAACCTCCCTGTTCTTATGGATTCGAACAGCATACCCCCGATTCCGCTACGACCAACTCATGCACCTCCTATGGAAAAACTTCCTACCACTCACCCTAGCATTACTTATGTGGTATGTCTCCATGCCCATTACAATCTCCAGCATTCCCCCTCAAACCTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGGTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCAGCCCCTGTCCTCCCTTCACCCCCATTGCGTATGAGCATTTCAGAACTCCAAGGAGTCACAGGCATCTTTATAGTTCACGTTAACATATAGACACTGTTGGAAGCAGTTCCTTCTAAAAGGGTAGCCCTGGACTTAATACCAGCCGGATACCTCTGGCCCCCACCCCATTACTGTACCTCTGGAGTCACTACTGTGGGTCGCCACTCCTCTGCTACACAGCACGGCTTTTTCAAGGCTGTATTGAGAAGGGAAGTTAGGAAGAAGGGTGTGCTGGGCTAACCAGCCCACAGAGCTCACATTCCTGTCCCTTGGGTGAAAAATACATGTCCATCCTGATATCTCCTGAATTCAGAAATTAGCCTCCACATGTGCAATGGCTTTAAGAGCCAGAAGCAGGGTTCTGGGTTTTGGAAGTTACCTGTGGCCAGGTGTGGTCTCGGTTACCAAATACGGTTACCTGCAGCTTTCTGGGTCCTTTGTGCTCCCACGGGTCTACAGAGTCCCATCTGCCCAAAGGTCTTGAAGCTTGACAGGATGTTTTCGATTACTCAGTCTCCCAGGGCACTACTGGTCCGTAGGATTCGATTGGTCGGGGTAGGAGAGTTAAACAACATTTAAACAGAGTTCTCTCAAAAATGTCTAAAGGGATTGTAGGTAGATAACATCCAATCACTGTTTGCACTTATCTGAAATCTTCCCTCTTGGCTGCCCCCAGGTATTTACTGTGGAGAACATTGCATAGGAATGTCTGGAAAAAGCTTCTACAACTTGTTACAGCCTTCACATTTGTAGAAGCTTT 54opt_COX10*-ND1-3′UTR*ATGGCCGCCAGCCCCCACACCCTGAGCAGCCGCCTGCTGACCGGCTGCGTGGGCGGCAGCGTGTGGTACCTGGAGCGCCGCACCATGGCCAACCTCCTACTCCTCATTGTACCCATTCTAATCGCAATGGCATTCCTAATGCTTACCGAACGAAAAATTCTAGGCTATATGCAACTACGCAAAGGCCCCAACGTTGTAGGCCCCTACGGGCTACTACAACCCTTCGCTGACGCCATAAAACTCTTCACCAAAGAGCCCCTAAAACCCGCCACATCTACCATCACCCTCTACATCACCGCCCCGACCTTAGCTCTCACCATCGCTCTTCTACTATGGACCCCCCTCCCCATGCCCAACCCCCTGGTCAACCTCAACCTAGGCCTCCTATTTATTCTAGCCACCTCTAGCCTAGCCGTTTACTCAATCCTCTGGTCAGGGTGGGCATCAAACTCAAACTACGCCCTGATCGGCGCACTGCGAGCAGTAGCCCAAACAATCTCATATGAAGTCACCCTAGCCATCATTCTACTATCAACATTACTAATGAGTGGCTCCTTTAACCTCTCCACCCTTATCACAACACAAGAACACCTCTGGTTACTCCTGCCATCATGGCCCTTGGCCATGATGTGGTTTATCTCCACACTAGCAGAGACCAACCGAACCCCCTTCGACCTTGCCGAAGGGGAGTCCGAACTAGTCTCAGGCTTCAACATCGAATACGCCGCAGGCCCCTTCGCCCTATTCTTCATGGCCGAATACACAAACATTATTATGATGAACACCCTCACCACTACAATCTTCCTAGGAACAACATATGACGCACTCTCCCCTGAACTCTACACAACATATTTTGTCACCAAGACCCTACTTCTAACCTCCCTGTTCTTATGGATTCGAACAGCATACCCCCGATTCCGCTACGACCAACTCATGCACCTCCTATGGAAAAACTTCCTACCACTCACCCTAGCATTACTTATGTGGTATGTCTCCATGCCCATTACAATCTCCAGCATTCCCCCTCAAACCTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGGTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCA 55 opt_COX10*-opt_ND1-3′UTRATGGCCGCCAGCCCCCACACCCTGAGCAGCCGCCTGCTGACCGGCTGCGTGGGCGGCAGCGTGTGGTACCTGGAGCGCCGCACCATGGCCAACCTGCTGCTGCTGATCGTGCCCATCCTGATCGCCATGGCCTTCCTGATGCTGACCGAGCGCAAGATCCTGGGCTACATGCAGCTGCGCAAGGGCCCCAACGTGGTGGGCCCCTACGGCCTGCTGCAGCCCTTCGCCGACGCCATCAAGCTGTTCACCAAGGAGCCCCTGAAGCCCGCCACCAGCACCATCACCCTGTACATCACCGCCCCCACCCTGGCCCTGACCATCGCCCTGCTGCTGTGGACCCCCCTGCCCATGCCCAACCCCCTGGTGAACCTGAACCTGGGCCTGCTGTTCATCCTGGCCACCAGCAGCCTGGCCGTGTACAGCATCCTGTGGAGCGGCTGGGCCAGCAACAGCAACTACGCCCTGATCGGCGCCCTGCGCGCCGTGGCCCAGACCATCAGCTACGAGGTGACCCTGGCCATCATCCTGCTGAGCACCCTGCTGATGAGCGGCAGCTTCAACCTGAGCACCCTGATCACCACCCAGGAGCACCTGTGGCTGCTGCTGCCCAGCTGGCCCCTGGCCATGATGTGGTTCATCAGCACCCTGGCCGAGACCAACCGCACCCCCTTCGACCTGGCCGAGGGCGAGAGCGAGCTGGTGAGCGGCTTCAACATCGAGTACGCCGCCGGCCCCTTCGCCCTGTTCTTCATGGCCGAGTACACCAACATCATCATGATGAACACCCTGACCACCACCATCTTCCTGGGCACCACCTACGACGCCCTGAGCCCCGAGCTGTACACCACCTACTTCGTGACCAAGACCCTGCTGCTGACCAGCCTGTTCCTGTGGATCCGCACCGCCTACCCCCGCTTCCGCTACGACCAGCTGATGCACCTGCTGTGGAAGAACTTCCTGCCCCTGACCCTGGCCCTGCTGATGTGGTACGTGAGCATGCCCATCACCATCAGCAGCATCCCCCCCCAGACCTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGGTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCAGCCCCTGTCCTCCCTTCACCCCCATTGCGTATGAGCATTTCAGAACTCCAAGGAGTCACAGGCATCTTTATAGTTCACGTTAACATATAGACACTGTTGGAAGCAGTTCCTTCTAAAAGGGTAGCCCTGGACTTAATACCAGCCGGATACCTCTGGCCCCCACCCCATTACTGTACCTCTGGAGTCACTACTGTGGGTCGCCACTCCTCTGCTACACAGCACGGCTTTTTCAAGGCTGTATTGAGAAGGGAAGTTAGGAAGAAGGGTGTGCTGGGCTAACCAGCCCACAGAGCTCACATTCCTGTCCCTTGGGTGAAAAATACATGTCCATCCTGATATCTCCTGAATTCAGAAATTAGCCTCCACATGTGCAATGGCTTTAAGAGCCAGAAGCAGGGTTCTGGGAATTTTGCAAGTTACCTGTGGCCAGGTGTGGTCTCGGTTACCAAATACGGTTACCTGCAGCTTTTTAGTCCTTTGTGCTCCCACGGGTCTACAGAGTCCCATCTGCCCAAAGGTCTTGAAGCTTGACAGGATGTTTTCGATTACTCAGTCTCCCAGGGCACTACTGGTCCGTAGGATTCGATTGGTCGGGGTAGGAGAGTTAAACAACATTTAAACAGAGTTCTCTCAAAAATGTCTAAAGGGATTGTAGGTAGATAACATCCAATCACTGTTTGCACTTATCTGAAATCTTCCCTCTTGGCTGCCCCCAGGTATTTACTGTGGAGAACATTGCATAGGAATGTCTGGAAAAAGCTTCTACAACTTGTTACAGCCTTCACATTTGTAGAAGCTTT 56 opt_COX10*-opt_ND1-3′UTR*ATGGCCGCCAGCCCCCACACCCTGAGCAGCCGCCTGCTGACCGGCTGCGTGGGCGGCAGCGTGTGGTACCTGGAGCGCCGCACCATGGCCAACCTGCTGCTGCTGATCGTGCCCATCCTGATCGCCATGGCCTTCCTGATGCTGACCGAGCGCAAGATCCTGGGCTACATGCAGCTGCGCAAGGGCCCCAACGTGGTGGGCCCCTACGGCCTGCTGCAGCCCTTCGCCGACGCCATCAAGCTGTTCACCAAGGAGCCCCTGAAGCCCGCCACCAGCACCATCACCCTGTACATCACCGCCCCCACCCTGGCCCTGACCATCGCCCTGCTGCTGTGGACCCCCCTGCCCATGCCCAACCCCCTGGTGAACCTGAACCTGGGCCTGCTGTTCATCCTGGCCACCAGCAGCCTGGCCGTGTACAGCATCCTGTGGAGCGGCTGGGCCAGCAACAGCAACTACGCCCTGATCGGCGCCCTGCGCGCCGTGGCCCAGACCATCAGCTACGAGGTGACCCTGGCCATCATCCTGCTGAGCACCCTGCTGATGAGCGGCAGCTTCAACCTGAGCACCCTGATCACCACCCAGGAGCACCTGTGGCTGCTGCTGCCCAGCTGGCCCCTGGCCATGATGTGGTTCATCAGCACCCTGGCCGAGACCAACCGCACCCCCTTCGACCTGGCCGAGGGCGAGAGCGAGCTGGTGAGCGGCTTCAACATCGAGTACGCCGCCGGCCCCTTCGCCCTGTTCTTCATGGCCGAGTACACCAACATCATCATGATGAACACCCTGACCACCACCATCTTCCTGGGCACCACCTACGACGCCCTGAGCCCCGAGCTGTACACCACCTACTTCGTGACCAAGACCCTGCTGCTGACCAGCCTGTTCCTGTGGATCCGCACCGCCTACCCCCGCTTCCGCTACGACCAGCTGATGCACCTGCTGTGGAAGAACTTCCTGCCCCTGACCCTGGCCCTGCTGATGTGGTACGTGAGCATGCCCATCACCATCAGCAGCATCCCCCCCCAGACCTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCA 57 COX8-ND4-3′UTRATGTCCGTCCTGACGCGCCTGCTGCTGCGGGGCTTGACACGGCTCGGCTCGGCGGCTCCAGTGCGGCGCGCCAGAATCCATTCGTTGATGCTAAAACTAATCGTCCCAACAATTATGTTACTACCACTGACATGGCTTTCCAAAAAACACATGATTTGGATCAACACAACCACCCACAGCCTAATTATTAGCATCATCCCTCTACTATTTTTTAACCAAATCAACAACAACCTATTTAGCTGTTCCCCAACCTTTTCCTCCGACCCCCTAACAACCCCCCTCCTAATGCTAACTACCTGGCTCCTACCCCTCACAATCATGGCAAGCCAACGCCACTTATCCAGTGAACCACTATCACGAAAAAAACTCTACCTCTCTATGCTAATCTCCCTACAAATCTCCTTAATTATGACATTCACAGCCACAGAACTAATCATGTTTTATATCTTCTTCGAAACCACACTTATCCCCACCTTGGCTATCATCACCCGATGGGGCAACCAGCCAGAACGCCTGAACGCAGGCACATACTTCCTATTCTACACCCTAGTAGGCTCCCTTCCCCTACTCATCGCACTAATTTACACTCACAACACCCTAGGCTCACTAAACATTCTACTACTCACTCTCACTGCCCAAGAACTATCAAACTCCTGGGCCAACAACTTAATGTGGCTAGCTTACACAATGGCTTTTATGGTAAAGATGCCTCTTTACGGACTCCACTTATGGCTCCCTAAAGCCCATGTCGAAGCCCCCATCGCTGGGTCAATGGTACTTGCCGCAGTACTCTTAAAACTAGGCGGCTATGGTATGATGCGCCTCACACTCATTCTCAACCCCCTGACAAAACACATGGCCTACCCCTTCCTTGTACTATCCCTATGGGGCATGATTATGACAAGCTCCATCTGCCTACGACAAACAGACCTAAAATCGCTCATTGCATACTCTTCAATCAGCCACATGGCCCTCGTAGTAACAGCCATTCTCATCCAAACCCCCTGGAGCTTCACCGGCGCAGTCATTCTCATGATCGCCCACGGGCTTACATCCTCATTACTATTCTGCCTAGCAAACTCAAACTACGAACGCACTCACAGTCGCATCATGATCCTCTCTCAAGGACTTCAAACTCTACTCCCACTAATGGCTTTTTGGTGGCTTCTAGCAAGCCTCGCTAACCTCGCCTTACCCCCCACTATTAACCTACTGGGAGAACTCTCTGTGCTAGTAACCACGTTCTCCTGGTCAAATATCACTCTCCTACTTACAGGACTCAACATGCTAGTCACAGCCCTATACTCCCTCTACATGTTTACCACAACACAATGGGGCTCACTCACCCACCACATTAACAACATGAAACCCTCATTCACACGAGAAAACACCCTCATGTTCATGCACCTATCCCCCATTCTCCTCCTATCCCTCAACCCCGACATCATTACCGGGTTTTCCTCTTAAGAGCACTGGGACGCCCACCGCCCCTTTcoCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCAGCCCCTGTCCTCCCTTCACCCCCATTGCGTATGAGCATTTCAGAACTCCAAGGAGTCACAGGCATCTTTATAGTTCACGTTAACATATAGACACTGTTGGAAGCAGTTCCTTCTAAAAGGGTAGCCCTGGACTTAATACCAGCCGGATACCTCTGGCCCCCACCCCATTACTGTACCTCTGGAGTCACTACTGTGGGTCGCCACTCCTCTGCTACACAGCACGGCTTTTTCAAGGCTGTATTGAGAAGGGAAGTTAGGAAGAAGGGTGTGCTGGGCTAACCAGCCCACAGAGCTCACATTCCTGTCCCTTGGGTGAAAAATACATGTCCATCCTGATATCTCCTGAATTCAGAAATTAGCCTCCACATGTGCAATGGCTTTAAGAGCCAGAAGCAGGGTTCTGGGAATTTTGCAAGTTACCTGTGGCCAGGTGTGGTCTCGGTTACCAAATACGGTTACCTGCAGCTTTTTAGTCCTTTGTGCTCCCACGGGTCTACAGAGTCCCATCTGCCCAAAGGTCTTGAAGCTTGACAGGATGTTTTCGATTACTCAGTCTCCCAGGGCACTACTGGTCCGTAGGATTCGATTGGTCGGGGTAGGAGAGTTAAACAACATTTAAACAGAGTTCTCTCAAAAATGTCTAAAGGGATTGTAGGTAGATAACATCCAATCACTGTTTGCACTTATCTGAAATCTTCCCTCTTGGCTGCCCCCAGGTATTTACTGTGGAGAACATTGCATAGGAATGTCTGGAAAAAGCTTCTACAACTTGTTACAGCCTTCACATTTGTAGAAGCTTT 58 COX8-ND4-3′UTR*ATGTCCGTCCTGACGCGCCTGCTGCTGCGGGGCTTGACACGGCTCGGCTCGGCGGCTCCAGTGCGGCGCGCCAGAATCCATTCGTTGATGCTAAAACTAATCGTCCCAACAATTATGTTACTACCACTGACATGGCTTTCCAAAAAACACATGATTTGGATCAACACAACCACCCACAGCCTAATTATTAGCATCATCCCTCTACTATTTTTTAACCAAATCAACAACAACCTATTTAGCTGTTCCCCAACCTTTTCCTCCGACCCCCTAACAACCCCCCTCCTAATGCTAACTACCTGGCTCCTACCCCTCACAATCATGGCAAGCCAACGCCACTTATCCAGTGAACCACTATCACGAAAAAAACTCTACCTCTCTATGCTAATCTCCCTACAAATCTCCTTAATTATGACATTCACAGCCACAGAACTAATCATGTTTTATATCTTCTTCGAAACCACACTTATCCCCACCTTGGCTATCATCACCCGATGGGGCAACCAGCCAGAACGCCTGAACGCAGGCACATACTTCCTATTCTACACCCTAGTAGGCTCCCTTCCCCTACTCATCGCACTAATTTACACTCACAACACCCTAGGCTCACTAAACATTCTACTACTCACTCTCACTGCCCAAGAACTATCAAACTCCTGGGCCAACAACTTAATGTGGCTAGCTTACACAATGGCTTTTATGGTAAAGATGCCTCTTTACGGACTCCACTTATGGCTCCCTAAAGCCCATGTCGAAGCCCCCATCGCTGGGTCAATGGTACTTGCCGCAGTACTCTTAAAACTAGGCGGCTATGGTATGATGCGCCTCACACTCATTCTCAACCCCCTGACAAAACACATGGCCTACCCCTTCCTTGTACTATCCCTATGGGGCATGATTATGACAAGCTCCATCTGCCTACGACAAACAGACCTAAAATCGCTCATTGCATACTCTTCAATCAGCCACATGGCCCTCGTAGTAACAGCCATTCTCATCCAAACCCCCTGGAGCTTCACCGGCGCAGTCATTCTCATGATCGCCCACGGGCTTACATCCTCATTACTATTCTGCCTAGCAAACTCAAACTACGAACGCACTCACAGTCGCATCATGATCCTCTCTCAAGGACTTCAAACTCTACTCCCACTAATGGCTTTTTGGTGGCTTCTAGCAAGCCTCGCTAACCTCGCCTTACCCCCCACTATTAACCTACTGGGAGAACTCTCTGTGCTAGTAACCACGTTCTCCTGGTCAAATATCACTCTCCTACTTACAGGACTCAACATGCTAGTCACAGCCCTATACTCCCTCTACATGTTTACCACAACACAATGGGGCTCACTCACCCACCACATTAACAACATGAAACCCTCATTCACACGAGAAAACACCCTCATGTTCATGCACCTATCCCCCATTCTCCTCCTATCCCTCAACCCCGACATCATTACCGGGTTTTCCTCTTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCA 59 COX8-opt_ND4-3′UTRATGTCCGTCCTGACGCGCCTGCTGCTGCGGGGCTTGACACGGCTCGGCTCGGCGGCTCCAGTGCGGCGCGCCAGAATCCATTCGTTGATGCTGAAGCTGATCGTGCCCACCATCATGCTGCTGCCTCTGACCTGGCTGAGCAAGAAACACATGATCTGGATCAACACCACCACGCACAGCCTGATCATCAGCATCATCCCTCTGCTGTTCTTCAACCAGATCAACAACAACCTGTTCAGCTGCAGCCCCACCTTCAGCAGCGACCCTCTGACAACACCTCTGCTGATGCTGACCACCTGGCTGCTGCCCCTCACAATCATGGCCTCTCAGAGACACCTGAGCAGCGAGCCCCTGAGCCGGAAGAAACTGTACCTGAGCATGCTGATCTCCCTGCAGATCTCTCTGATCATGACCTTCACCGCCACCGAGCTGATCATGTTCTACATCTTTTTCGAGACAACGCTGATCCCCACACTGGCCATCATCACCAGATGGGGCAACCAGCCTGAGAGACTGAACGCCGGCACCTACTTTCTGTTCTACACCCTCGTGGGCAGCCTGCCACTGCTGATTGCCCTGATCTACACCCACAACACCCTGGGCTCCCTGAACATCCTGCTGCTGACACTGACAGCCCAAGAGCTGAGCAACAGCTGGGCCAACAATCTGATGTGGCTGGCCTACACAATGGCCTTCATGGTCAAGATGCCCCTGTACGGCCTGCACCTGTGGCTGCCTAAAGCTCATGTGGAAGCCCCTATCGCCGGCTCTATGGTGCTGGCTGCAGTGCTGCTGAAACTCGGCGGCTACGGCATGATGCGGCTGACCCTGATTCTGAATCCCCTGACCAAGCACATGGCCTATCCATTTCTGGTGCTGAGCCTGTGGGGCATGATTATGACCAGCAGCATCTGCCTGCGGCAGACCGATCTGAAGTCCCTGATCGCCTACAGCTCCATCAGCCACATGGCCCTGGTGGTCACCGCCATCCTGATTCAGACCCCTTGGAGCTTTACAGGCGCCGTGATCCTGATGATTGCCCACGGCCTGACAAGCAGCCTGCTGTTTTGTCTGGCCAACAGCAACTACGAGCGGACCCACAGCAGAATCATGATCCTGTCTCAGGGCCTGCAGACCCTCCTGCCTCTTATGGCTTTTTGGTGGCTGCTGGCCTCTCTGGCCAATCTGGCACTGCCTCCTACCATCAATCTGCTGGGCGAGCTGAGCGTGCTGGTCACCACATTCAGCTGGTCCAATATCACCCTGCTGCTCACCGGCCTGAACATGCTGGTTACAGCCCTGTACTCCCTGTACATGTTCACCACCACACAGTGGGGAAGCCTGACACACCACATCAACAATATGAAGCCCAGCTTCACCCGCGAGAACACCCTGATGTTCATGCATCTGAGCCCCATTCTGCTGCTGTCCCTGAATCCTGATATCATCACCGGCTTCTCCAGCTGAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCAGCCCCTGTCCTCCCTTCACCCCCATTGCGTATGAGCATTTCAGAACTCCAAGGAGTCACAGGCATCTTTATAGTTCACGTTAACATATAGACACTGTTGGAAGCAGTTCCTTCTAAAAGGGTAGCCCTGGACTTAATACCAGCCGGATACCTCTGGCCCCCACCCCATTACTGTACCTCTGGAGTCACTACTGTGGGTCGCCACTCCTCTGCTACACAGCACGGCTTTTTCAAGGCTGTATTGAGAAGGGAAGTTAGGAAGAAGGGTGTGCTGGGCTAACCAGCCCACAGAGCTCACATTCCTGTCCCTTGGGTGAAAAATACATGTCCATCCTGATATCTCCTGAATTCAGAAATTAGCCTCCACATGTGCAATGGCTTTAAGAGCCAGAAGCAGGGTTCTGGGAATTTTGCAAGTTACCTGTGGCCAGGTGTGGTCTCGGTTACCAAATACGGTTACCTGCAGCTTTTTAGTCCTTTGTGCTCCCACGGGTCTACAGAGTCCCATCTGCCCAAAGGTCTTGAAGCTTGACAGGATGTTTTCGATTACTCAGTCTCCCAGGGCACTACTGGTCCGTAGGATTCGATTGGTCGGGGTAGGAGAGTTAAACAACATTTAAACAGAGTTCTCTCAAAAATGTCTAAAGGGATTGTAGGTAGATAACATCCAATCACTGTTTGCACTTATCTGAAATCTTCCCTCTTGGCTGCCCCCAGGTATTTACTGTGGAGAACATTGCATAGGAATGTCTGGAAAAAGCTTCTACAACTTGTTACAGCCTTCACATTTGTAGAAGCTTT 60 COX8-opt_ND4-3′UTR*ATGTCCGTCCTGACGCGCCTGCTGCTGCGGGGCTTGACACGGCTCGGCTCGGCGGCTCCAGTGCGGCGCGCCAGAATCCATTCGTTGATGCTGAAGCTGATCGTGCCCACCATCATGCTGCTGCCTCTGACCTGGCTGAGCAAGAAACACATGATCTGGATCAACACCACCACGCACAGCCTGATCATCAGCATCATCCCTCTGCTGTTCTTCAACCAGATCAACAACAACCTGTTCAGCTGCAGCCCCACCTTCAGCAGCGACCCTCTGACAACACCTCTGCTGATGCTGACCACCTGGCTGCTGCCCCTCACAATCATGGCCTCTCAGAGACACCTGAGCAGCGAGCCCCTGAGCCGGAAGAAACTGTACCTGAGCATGCTGATCTCCCTGCAGATCTCTCTGATCATGACCTTCACCGCCACCGAGCTGATCATGTTCTACATCTTTTTCGAGACAACGCTGATCCCCACACTGGCCATCATCACCAGATGGGGCAACCAGCCTGAGAGACTGAACGCCGGCACCTACTTTCTGTTCTACACCCTCGTGGGCAGCCTGCCACTGCTGATTGCCCTGATCTACACCCACAACACCCTGGGCTCCCTGAACATCCTGCTGCTGACACTGACAGCCCAAGAGCTGAGCAACAGCTGGGCCAACAATCTGATGTGGCTGGCCTACACAATGGCCTTCATGGTCAAGATGCCCCTGTACGGCCTGCACCTGTGGCTGCCTAAAGCTCATGTGGAAGCCCCTATCGCCGGCTCTATGGTGCTGGCTGCAGTGCTGCTGAAACTCGGCGGCTACGGCATGATGCGGCTGACCCTGATTCTGAATCCCCTGACCAAGCACATGGCCTATCCATTTCTGGTGCTGAGCCTGTGGGGCATGATTATGACCAGCAGCATCTGCCTGCGGCAGACCGATCTGAAGTCCCTGATCGCCTACAGCTCCATCAGCCACATGGCCCTGGTGGTCACCGCCATCCTGATTCAGACCCCTTGGAGCTTTACAGGCGCCGTGATCCTGATGATTGCCCACGGCCTGACAAGCAGCCTGCTGTTTTGTCTGGCCAACAGCAACTACGAGCGGACCCACAGCAGAATCATGATCCTGTCTCAGGGCCTGCAGACCCTCCTGCCTCTTATGGCTTTTTGGTGGCTGCTGGCCTCTCTGGCCAATCTGGCACTGCCTCCTACCATCAATCTGCTGGGCGAGCTGAGCGTGCTGGTCACCACATTCAGCTGGTCCAATATCACCCTGCTGCTCACCGGCCTGAACATGCTGGTTACAGCCCTGTACTCCCTGTACATGTTCACCACCACACAGTGGGGAAGCCTGACACACCACATCAACAATATGAAGCCCAGCTTCACCCGCGAGAACACCCTGATGTTCATGCATCTGAGCCCCATTCTGCTGCTGTCCCTGAATCCTGATATCATCACCGGCTTCTCCAGCTGAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCA 61COX8-opt_ND4*-3′UTRATGTCCGTCCTGACGCGCCTGCTGCTGCGGGGCTTGACACGGCTCGGCTCGGCGGCTCCAGTGCGGCGCGCCAGAATCCATTCGTTGATGCTGAAGCTGATCGTGCCCACCATCATGCTGCTGCCCCTGACCTGGCTGAGCAAGAAGCACATGATCTGGATCAACACCACCACCCACAGCCTGATCATCAGCATCATCCCCCTGCTGTTCTTCAACCAGATCAACAACAACCTGTTCAGCTGCAGCCCCACCTTCAGCAGCGACCCCCTGACCACCCCCCTGCTGATGCTGACCACCTGGCTGCTGCCCCTGACCATCATGGCCAGCCAGCGCCACCTGAGCAGCGAGCCCCTGAGCCGCAAGAAGCTGTACCTGAGCATGCTGATCAGCCTGCAGATCAGCCTGATCATGACCTTCACCGCCACCGAGCTGATCATGTTCTACATCTTCTTCGAGACCACCCTGATCCCCACCCTGGCCATCATCACCCGCTGGGGCAACCAGCCCGAGCGCCTGAACGCCGGCACCTACTTCCTGTTCTACACCCTGGTGGGCAGCCTGCCCCTGCTGATCGCCCTGATCTACACCCACAACACCCTGGGCAGCCTGAACATCCTGCTGCTGACCCTGACCGCCCAGGAGCTGAGCAACAGCTGGGCCAACAACCTGATGTGGCTGGCCTACACCATGGCCTTCATGGTGAAGATGCCCCTGTACGGCCTGCACCTGTGGCTGCCCAAGGCCCACGTGGAGGCCCCCATCGCCGGCAGCATGGTGCTGGCCGCCGTGCTGCTGAAGCTGGGCGGCTACGGCATGATGCGCCTGACCCTGATCCTGAACCCCCTGACCAAGCACATGGCCTACCCCTTCCTGGTGCTGAGCCTGTGGGGCATGATCATGACCAGCAGCATCTGCCTGCGCCAGACCGACCTGAAGAGCCTGATCGCCTACAGCAGCATCAGCCACATGGCCCTGGTGGTGACCGCCATCCTGATCCAGACCCCCTGGAGCTTCACCGGCGCCGTGATCCTGATGATCGCCCACGGCCTGACCAGCAGCCTGCTGTTCTGCCTGGCCAACAGCAACTACGAGCGCACCCACAGCCGCATCATGATCCTGAGCCAGGGCCTGCAGACCCTGCTGCCCCTGATGGCCTTCTGGTGGCTGCTGGCCAGCCTGGCCAACCTGGCCCTGCCCCCCACCATCAACCTGCTGGGCGAGCTGAGCGTGCTGGTGACCACCTTCAGCTGGAGCAACATCACCCTGCTGCTGACCGGCCTGAACATGCTGGTGACCGCCCTGTACAGCCTGTACATGTTCACCACCACCCAGTGGGGCAGCCTGACCCACCACATCAACAACATGAAGCCCAGCTTCACCCGCGAGAACACCCTGATGTTCATGCACCTGAGCCCCATCCTGGTGCTGAGCCTGAACCCCGACATCATCACCGGCTTCAGCAGCTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCAGCCCCTGTCCTCCCTTCACCCCCATTGCGTATGAGCATTTCAGAACTCCAAGGAGTCACAGGCATCTTTATAGTTCACGTTAACATATAGACACTGTTGGAAGCAGTTCCTTCTAAAAGGGTAGCCCTGGACTTAATACCAGCCGGATACCTCTGGCCCCCACCCCATTACTGTACCTCTGGAGTCACTACTGTGGGTCGCCACTCCTCTGCTACACAGCACGGCTTTTTCAAGGCTGTATTGAGAAGGGAAGTTAGGAAGAAGGGTGTGCTGGGCTAACCAGCCCACAGAGCTCACATTCCTGTCCCTTGGGTGAAAAATACATGTCCATCCTGATATCTCCTGAATTCAGAAATTAGCCTCCACATGTGCAATGGCTTTAAGAGCCAGAAGCAGGGTTCTGGGAATTTTGCAAGTTACCTGTGGCCAGGTGTGGTCTCGGTTACCAAATACGGTTACCTGCAGCTTTTTAGTCCTTTGTGCTCCCACGGGTCTACAGAGTCCCAlCTGCCCAAAGGTCTTGAAGCTTGACAGGATGTTTTCGATTACTCAGTCTCCCAGGGCACTACTGGTCCGTAGGATTCGATTGGTCGGGGTAGGAGAGTTAAACAACATTTAAACAGAGTTCTCTCAAAAATGTCTAAAGGGATTGTAGGTAGATAACATCCAATCACTGTTTGCACTTATCTGAAATCTTCCCTCTTGGCTGCCCCCAGGTATTTACTGTGGAGAACATTGCATAGGAATGTCTGGAAAAAGCTTCTACAACTTGTTACAGCCTGTCACATTTTAGAAGCTTT 62COX8-opt_ND4*-3′UTR*ATGTCCGTCCTGACGCGCCTGCTGCTGCGGGGCTTGACACGGCTCGGCTCGGCGGCTCCAGTGCGGCGCGCCAGAATCCATTCGTTGATGCTGAAGCTGATCGTGCCCACCATCATGCTGCTGCCCCTGACCTGGCTGAGCAAGAAGCACATGATCTGGATCAACACCACCACCCACAGCCTGATCATCAGCATCATCCCCCTGCTGTTCTTCAACCAGATCAACAACAACCTGTTCAGCTGCAGCCCCACCTTCAGCAGCGACCCCCTGACCACCCCCCTGCTGATGCTGACCACCTGGCTGCTGCCCCTGACCATCATGGCCAGCCAGCGCCACCTGAGCAGCGAGCCCCTGAGCCGCAAGAAGCTGTACCTGAGCATGCTGATCAGCCTGCAGATCAGCCTGATCATGACCTTCACCGCCACCGAGCTGATCATGTTCTACATCTTCTTCGAGACCACCCTGATCCCCACCCTGGCCATCATCACCCGCTGGGGCAACCAGCCCGAGCGCCTGAACGCCGGCACCTACTTCCTGTTCTACACCCTGGTGGGCAGCCTGCCCCTGCTGATCGCCCTGATCTACACCCACAACACCCTGGGCAGCCTGAACATCCTGCTGCTGACCCTGACCGCCCAGGAGCTGAGCAACAGCTGGGCCAACAACCTGATGTGGCTGGCCTACACCATGGCCTTCATGGTGAAGATGCCCCTGTACGGCCTGCACCTGTGGCTGCCCAAGGCCCACGTGGAGGCCCCCATCGCCGGCAGCATGGTGCTGGCCGCCGTGCTGCTGAAGCTGGGCGGCTACGGCATGATGCGCCTGACCCTGATCCTGAACCCCCTGACCAAGCACATGGCCTACCCCTTCCTGGTGCTGAGCCTGTGGGGCATGATCATGACCAGCAGCATCTGCCTGCGCCAGACCGACCTGAAGAGCCTGATCGCCTACAGCAGCATCAGCCACATGGCCCTGGTGGTGACCGCCATCCTGATCCAGACCCCCTGGAGCTTCACCGGCGCCGTGATCCTGATGATCGCCCACGGCCTGACCAGCAGCCTGCTGTTCTGCCTGGCCAACAGCAACTACGAGCGCACCCACAGCCGCATCATGATCCTGAGCCAGGGCCTGCAGACCCTGCTGCCCCTGATGGCCTTCTGGTGGCTGCTGGCCAGCCTGGCCAACCTGGCCCTGCCCCCCACCATCAACCTGCTGGGCGAGCTGAGCGTGCTGGTGACCACCTTCAGGTGGAGCAACATCACCCTGCTGCTGACCGGCCTGAACATGCTGGTGACCGCCCTGTACAGCCTGTACATGTTCACCACCACCCAGTGGGGCAGCCTGACCCACCACATCAACAACATGAAGCCCAGCTTCACCCGCGAGAACACCCTGATGTTCATGCACCTGAGCCCCATCCTGCTGCTGAGCCTGAACCCCGACATCATCACCGGCTTCAGCAGCTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTG CCA63 COX8-ND6-3′UTRATGTCCGTCCTGACGCGCCTGCTGCTGCGGGGCTTGACACGGCTCGGCTCGGCGGCTCCAGTGCGGCGCGCCAGAATCCATTCGTTGATGATGTATGCTTTGTTTCTGTTGAGTGTGGGTTTAGTAATGGGGTTTGTGGGGTTTTCTTCTAAGCCTTCTCCTATTTATGGGGGTTTAGTATTGATTGTTAGCGGTGTGGTCGGGTGTGTTATTATTCTGAATTTTGGGGGAGGTTATATGGGTTTAATGGTTTTTTTAATTTATTTAGGGGGAATGATGGTTGTCTTTGGATATACTACAGCGATGGCTATTGAGGAGTATCCTGAGGCATGGGGGTCAGGGGTTGAGGTCTTGGTGAGTGTTTTAGTGGGGTTAGCGATGGAGGTAGGATTGGTGCTGTGGGTGAAAGAGTATGATGGGGTGGTGGTTGTGGTAAACTTTAATAGTGTAGGAAGCTGGATGATTTATGAAGGAGAGGGGTCAGGGTTGATTCGGGAGGATCCTATTGGTGCGGGGGCTTTGTATGATTATGGGCGTTGGTTAGTAGTAGTTACTGGTTGGACATTGTTTGTTGGTGTATATATTGTAATTGAGATTGCTCGGGGGAATTAGGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCAGCCCCTGTCCTCCCTTCACCCCCATTGCGTATGAGCATTTCAGAACTCCAAGGAGTCACAGGCATCTTTATAGTTCACGTTAACATATAGACACTGTTGGAAGCAGTTCCTTCTAAAAGGGTAGCCCTGGACTTAATACCAGCCGGATACCTCTGGCCCCCACCCCATTACTGTACCTCTGGAGTCACTACTGTGGGTCGCCACTCCTCTGCTACACAGCACGGCTTTTTCAAGGCTGTATTGAGAAGGGAAGTTAGGAAGAAGGGTGTGCTGGGCTAACCAGCCCACAGAGCTCACATTCCTGTCCCTTGGGTGAAAAATACATGTCCATCCTGATATCTCCTGAATTCAGAAATTAGCCTCCACATGTGCAATGGCTTTAAGAGCCAGAAGCAGGGTTCTGGGAATTTTGCAAGTTACCTGTGGCCAGGTGTGGTCTCGGTTACCAAATACGGTTACCTGCAGCTTTTTAGTCCTTTGTGCTCCCACGGGTCTACAGAGTCCCATCTGCCCAAAGGTCTTGAAGCTTGACAGGATGTTTTCGATTACTCAGTCTCCCAGGGCACTACTGGTCCGTAGGATTCGATTGGTCGGGGTAGGAGAGTTAAACAACATTTAAACAGAGTTCTCTCAAAAATGTCTAAAGGGATTGTAGGTAGATAACATCCAATCACTGTTTGCACTTATCTGAAATCTTCCCTCTTGGCTGCCCCCAGGTATTTACTGTGGAGAACATTGCATAGGAATGTCTGGAAAAAGCTTCTACAACTTGTTACAGCCTTCACATTTGTAGAAGCTTT 64COX8-ND6-3′UTR*ATGTCCGTCCTGACGCGCCTGCTGCTGCGGGGCTTGACACGGCTCGGCTCGGCGGCTCCAGTGCGGCGCGCCAGAATCCATTCGTTGATGATGTATGCTTTGTTTCTGTTGAGTGTGGGTTTAGTAATGGGGTTTGTGGGGTTTTCTTCTAAGCCTTCTCCTATTTATGGGGGTTTAGTATTGATTGTTAGCGGTGTGGTCGGGTGTGTTATTATTCTGAATTTTGGGGGAGGTTATATGGGTTTAATGGTTTTTTTAATTTATTTAGGGGGAATGATGGTTGTCTTTGGATATACTACAGCGATGGCTATTGAGGAGTATCCTGAGGCATGGGGGTCAGGGGTTGAGGTCTTGGTGAGTGTTTTAGTGGGGTTAGCGATGGAGGTAGGATTGGTGCTGTGGGTGAAAGAGTATGATGGGGTGGTGGTTGTGGTAAACTTTAATAGTGTAGGAAGCTGGATGATTTATGAAGGAGAGGGGTCAGGGTTGATTCGGGAGGATCCTATTGGTGCGGGGGCTTTGTATGATTATGGGCGTTGGTTAGTAGTAGTTACTGGTTGGACATTGTTTGTTGGTGTATATATTGTAATTGAGATTGCTCGGGGGAATTAGGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCA 65 COX8-opt_ND6-3′UTRATGTCCGTCCTGACGCGCCTGCTGCTGCGGGGCTTGACACGGCTCGGCTCGGCGGCTCCAGTGCGGCGCGCCAGAATCCATTCGTTGATGATGTACGCCCTGTTCCTGCTGAGCGTGGGCCTGGTGATGGGCTTCGTGGGCTTCAGCAGCAAGCCCAGCCCCATCTACGGCGGCCTGGTGCTGATCGTGAGCGGCGTGGTGGGCTGCGTGATCATCCTGAACTTCGGCGGCGGCTACATGGGCCTGATGGTGTTCCTGATCTACCTGGGCGGCATGATGGTGGTGTTCGGCTACACCACCGCCATGGCCATCGAGGAGTACCCCGAGGCCTGGGGCAGCGGCGTGGAGGTGCTGGTGAGCGTGCTGGTGGGCCTGGCCATGGAGGTGGGCCTGGTGCTGTGGGTGAAGGAGTACGACGGCGTGGTGGTGGTGGTGAACTTCAACAGCGTGGGCAGCTGGATGATCTACGAGGGCGAGGGCAGCGGCCTGATCCGCGAGGACCCCATCGGCGCCGGCGCCCTGTACGACFACGGCCGCTGGCTGGTGGTGGTGACCGGCTGGACCCTGTTCGTGGGCGTGTACATCGTGATCGAGATCGCCCGCGGCAACTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTTATTTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCAGCCCCTGTCCTCCCTTCACCCCCATTGCGTATGAGCATTTCAGAACTCCAAGGAGTCACAGGCATCTTTATAGTTCACGTTAACATATAGACACTGTTGGAAGCAGTTCCTTCTAAAAGGGTAGCCCTGGACTTAATACCAGCCGGATACCTCTGGCCCCCACCCCATTACTGTACCTCTGGAGTCACTACTGTGGGTCGCCACTCCTCTGCTACACAGCACGGCTTTTTCAAGGCTGTATTGAGAAGGGAAGTTAGGAAGAAGGGTGTGCTGGGCTAACCAGCCCACAGAGCTCACATTCCTGTCCCTTGGGTGAAAAATACATGTCCATCCTGATATCTCCTGAATTCAGAAATTAGCCTCCACATGTGCAATGGCTTTAAGAGCCAGAAGCAGGGTTCTGGGAATTTTGCAAGTTACCTGTGGCCAGGTGTGGTCTCGGTTACCAAATACGGTTACCTGCAGCTTTTTAGTCCTTTGTGCTCCCACGGGTCTACAGAGTCCCATCTGCCCAAAGGTCTTGAAGCTTGACAGGATGTTTTCGATTACTCAGTCTCCCAGGGCACTACTGGTCCGTAGGATTCGATTGGTCGGGGTAGGAGAGTTAAACAACATTTAAACAGAGTTCTCTCAAAAATGTCTAAAGGGATTGTAGGTAGATAACATCCAATCACTGTTTGCACTTATCTGAAATCTTCCCTCTTGGCTGCCCCCAGGTATTTACTGTGGAGAACATTGCATAGGAATGTCTGGAAAAAAGCTTCTACAACTTGTTACAGCCTTCACATTTGTAGAAGCTTT 66 COX8-opt_ND6-3′UTRATGTCCGTCCTGACGCGCCTGCTGCTGCGGGGCTTGACACGGCTCGGCTCGGCGGCTCCAGTGCGGCGCGCCAGAATCCATTCGTTGATGATGTACGCCCTGTTCCTGCTGAGCGTGGGCCTGGTGATGGGCTTCGTGGGCTTCAGCAGCAAGCCCAGCCCCATCTACGGCGGCCTGGTGCTGATCGTGAGCGGCGTGGTGGGCTGCGTGATCATCCTGAACTTCGGCGGCGGCTACATGGGCCTGATGGTGTTCCTGATCTACCTGGGCGGCATGATGGTGGTGTTCGGCTACACCACCGCCATGGCCATCGAGGAGTACCCCGAGGCCTGGGGCAGCGGCGTGGAGGTGCTGGTGAGCGTGCTGGTGGGCCTGGCCATGGAGGTGGGCCTGGTGCTGTGGGTGAAGGAGTACGACGGCGTGGTGGTGGTGGTGAACTTCAACAGCGTGGGCAGCTGGATGATCTACGAGGGCGAGGGCAGCGGCCTGATCCGCGAGGACCCCATCGGCGCCGGCGCCCTGTACGACTACGGCCGCTGGCTGGTGGTGGTGACCGGCTGGACCCTGTTCGTGGGCGTGTACATCGTGATCGAGATCGCCCGCGGCAACTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGATGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCA 67 COX8-ND1-3′UTRATGTCCGTCCTGACGCGCCTGCTGCTGCGGGGCTTGACACGGCTCGGCTCGGCGGCTCCAGTGCGGCGCGCCAGAATCCATTCGTTGATGGCCAACCTCCTACTCCTCATTGTACCCATTCTAATCGCAATGGCATTCCTAATGCTTACCGAACGAAAAATTCTAGGCTATATGCAACTACGCAAAGGCCCCAACGTTGTAGGCCCCTACGGGCTACTACAACCCTTCGCTGACGCCATAAAACTCTTCACCAAAGAGCCCCTAAAACCCGCCACATCTACCATCACCCTCTACATCACCGCCCCGACCTTAGCTCTCACCATCGCTCTTCTACTATGGACCCCCCTCCCCATGCCCAACCCCCTGGTCAACCTCAACCTAGGCCTCCTATTTATTCTAGCCACCTCTAGCCTAGCCGTTTACTCAATCCTCTGGTCAGGGTGGGCATCAAACTCAAACTACGCCCTGATCGGCGCACTGCGAGCAGTAGCCCAAACAATCTCATATGAAGTCACCCTAGCCATCATTCTACTATCAACATTACTAATGAGTGGCTCCTTTAACCTCTCCACCCTTATCACAACACAAGAACACCTCTGGTTACTCCTGCCATCATGGCCCTTGGCCATGATGTGGTTTATCTCCACACTAGCAGAGACCAACCGAACCCCCTTCGACCTTGCCGAAGGGGAGTCCGAACTAGTCTCAGGCTTCAACATCGAATACGCCGCAGGCCCCTTCGCCCTATTCTTCATGGCCGAATACACAAACATTATTATGATGAACACCCTCACCACTACAATCTTCCTAGGAACAACATATGACGCACTCTCCCCTGAACTCTACACAACATATTTTGTCACCAAGACCCTACTTCTAACCTCCCTGTTCTTATGGATTCGAACAGCATACCCCCGATTCCGCTACGACCAACTCATGCACCTCCTATGGAAAAACTTCCTACCACTCACCCTAGCATTACTTATGTGGTATGTCTCCATGCCCATTACAATCTCCAGCATTCCCCCTCAAACCTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCAGCCCCTGTCCTCCCTTCACCCCCATTGCGTATGAGCATTTCAGAACTCCAAGGAGTCACAGGCATCTTTATAGTTCACGTCATATAGACACTGTTGGAAGCAGTTCCTTCTAAAAGGGTAGCCCTGGACTTAATACCAGCCGGATACCTCTGGCCCCCACCCCATTACTGTACCTCTGGAGTCACTACTGTGGGTCGCCACTCCTCTGCTACACAGCACGGCTTTTTCAAGGCTGTATTGAGAAGGGAAGTTAGGAAGAAGGGTGTGCTGGGCTAACCAGCCCACAGAGCTCACATTCCTGTCCCTTGGGTGAAAAATACATGTCCATCCTGATATCTCCTGAATTCAGAAATTAGCCTCCACATGTGCAATGGCTTTAAGAGCCAGAAGCAGGGTTCTGGGAATTTTGCAAGTTACCTGTGGCCAGGTGTGGTCTCGGTTACCAAATACGGTTACCTGCAGCTTTTTAGTCCTTTGTGCTCCCACGGGTCTACAGAGTCCCATCTGCCCAAAGGTCTTGAAGCTTGACAGGATGTTTTCGATTACTCAGTCTCCCAGGGCACTACTGGTCCGTAGGATTCGATTGGTCGGGGTAGGAGAGTTAAACAACATTTAAACAGAGTTCTCTCAAAAATGTCTAAAGGGATTGTAGGTAGATAACATCCAATCACTGTTTGCACTTATCTGAAATCTTCCCTCTTGGCTGCCCCCAGGTATTTACTGTGGAGAACATTGCATAGGAATGTCTGGAAAAAGCTTCTACAACTTGTTACAGCCTTCACATTTGTAGAAGCTT T68 COX8-ND1-3′UTR*ATGTCCGTCCTGACGCGCCTGCTGCTGCGGGGCTTGACACGGCTCGGCTCGGCGGCTCCAGTGCGGCGCGCCAGAATCCATTCGTTGATGGCCAACCTCCTACTCCTCATTGTACCCATTCTAAATCGCAATGGCATTCCTAATGCTTACCGAACGAAAAATTCTAGGCTATATGCAACTACGCAAAGGCCCCAACGTTGTAGGCCCCTACGGGCTACTACAACCCTTCGCTGACGCCATAAAACTCTTCACCAAAGAGCCCCTAAAACCCGCCACATCTACCATCACCCTCTACATCACCGCCCCGACCTTAGCTCTCACCATCGCTCTTCTACTATGGACCCCCCTCCCCATGCCCAACCCCCTGGTCAACCTCAACCTAGGCCTCCTATTTATTCTAGCCACCTCTAGCCTAGCCGTTTACTCAATCCTCTGGTCAGGGTGGGCATCAAACTCAAACTACGCCCTGATCGGCGCACTGCGAGCAGTAGCCCAAACAATCTCATATGAAGTCACCCTAGCCATCATTCTACTATCAACATTACTAATGAGTGGCTCCTTTAACCTCTCCACCCTTATCACAACACAAGAACACCTCTGGTTACTCCTGCCATCATGGCCCTTGGCCATGATGTGGTTTATCTCCACACTAGCAGAGACCAACCGAACCCCCTTCGACCTTGCCGAAGGGGAGTCCGAACTAGTCTCAGGCTTCAAACATCGAATACGCCGCAGGCCCCTTCGCCCTATTCTTCATGGCCGAATACACAAACATTATTATGATGAACACCCTCACCACTACAATCTTCCTAGGAACAACATATGACGCACTCTCCCCTGAACTCTACACAACATATTTTGTCACCAAGACCCTACTTCTAACCTCCCTGTTCTTATGGATTCGAACAGCATACCCCCGATTCCGCTACGACCAACTCATGCACCTCCTATGGAAAAACTTCCTACCACTCACCCTAGCATTACTTATGTGGTATGTCTCCATGCCCATTACAATCTCCAGCATTCCCCCTCAAACCTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCA 69 COX8-opt_ND1-3′UTRATGTCCGTCCTGACGCGCCTGCTGCTGCGGGGCTTGACACGGCTCGGCTCGGCGGCTCCAGTGCGGCGCGCCAGAATCCATTCGTTGATGGCCAACCTGCTGCTGCTGATCGTGCCCATCCTGATCGCCATGGCCTTCCTGATGCTGACCGAGCGCAAGATCCTGGGCTACATGCAGCTGCGCAAGGGCCCCAACGTGGTGGGCCCCTACGGCCTGCTGCAGCCCTTCGCCGACGCCATCAAGCTGTTCACCAAGGAGCCCCTGAAGCCCGCCACCAGCACCATCACCCTGTACATCACCGCCCCCACCCTGGCCCTGACCATCGCCCTGCTGCTGTGGACCCCCCTGCCCATGCCCAACCCCCTGGTGAACCTGAACCTGGGCCTGCTGTTCATCCTGGCCACCAGCAGCCTGGCCGTGTACAGCATCCTGTGGAGCGGCTGGGCCAGCAACAGCAACTACGCCCTGATCGGCGCCCTGCGCGCCGTGGCCCAGACCATCAGCTACGAGGTGACCCTGGCCATCATCCTGCTGAGCACCCTGCTGATGAGCGGCAGCTTCAACCTGAGCACCCTGATCACCACCCAGGAGCACCTGTGGCTGCTGCTGCCCAGCTGGCCCCTGGCCATGATGTGGTTCATCAGCACCCTGGCCGAGACCAACCGCACCCCCTTCGACCTGGCCGAGGGCGAGAGCGAGCTGGTGAGCGGCTTCAACATCGAGTACGCCGCCGGCCCCTTCGCCCTGTTCTTCATGGCCGAGTACACCAACATCATCATGATGAACACCCTGACCACCACCATCTTCCTGGGCACCACCTACGACGCCCTGAGCCCCGAGCTGTACACCACCTACTTCGTGACCAAGACCCTGCTGCTGACCAGCCTGTTCCTGTGGATCCGCACCGCCTACCCCCGCTTCCGCTACGACCAGCTGATGCACCTGCTGTGGAAGAACTTCCTGCCCCTGACCCTGGCCCTGCTGATGTGGTACGTGAGCATGCCCATCACCATCAGCAGCATCCCCCCCCAGACCTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCAGCCCCTGTCCTCCCTTCACCCCCATTGCGTATGAGCATTTCAGAACTCCAAGGAGTCACAGGCATCTTTATAGTTCACGTTAACATATAGACACTGTTGGAAGCAGTTCCTTCTAAAAGGGTAGCCCTGGACTTAATACCAGCCGGATACCTCTGGCCCCCACCCCATTACTGTACCTCTGGAGTCACTACTGTGGGTCGCCACTCCTCTGCTACACAGCACGGCTTTTTCAAGGCTGTATTGAGAAGGGAAGTTAGGAAGAAGGGTGTGCTGGGCTAACCAGCCCACAGAGCTCACATTCCTGTCCCTTGGGTGAAAAATACATGTCCATCCTGATATCTCCTGAATTCAGAAATTAGCCTCCACATGTGCAATGGCTTTAAGAGCCAGAAGCAGGGTTCTGGGAATTTTGCAAGTTACCTGTGGCCAGGTGTGGTCTCGGTTACCAAATACGGTTACCTGCAGCTTTTTAGTCCTTTGTGCTCCCACGGGTCTACAGAGTCCCATCTGCCCAAAGGTCTTGAAGCTTGACAGGATGTTTTCGATTACTCAGTCTCCCAGGGCACTACTGGTCCGTAGGATTCGATTGGTCGGGGTAGGAGAGTTAAACAACATTTAAACAGAGTTCTCTCAAAAATGTCTAAAGGGATTGTAGGTAGATAACATCCAATCACTGTTTGCACTTATCTGAAATCTTCCCTCTTGGCTGCCCCCAGGTATTTACTGTGGAGAACATTGCATAGGAATGTCTGGAAAAAGCTTCTACAAACTTGTTACAGCCTTCACATTTGTAGAAGCTTT 70 COX8-opt_ND1-3′UTR*ATGTCCGTCCTGACGCGCCTGCTGCTGCGGGGCTTGACACGGCTCGGCTCGGCGGCTCCAGTGCGGCGCGCCAGAATCCATTCGTTGATGGCCAACCTGCTGCTGCTGATCGTGCCCATCCTGATCGCCATGGCCTTCCTGATGCTGACCGAGCGCAAGATCCTGGGCTACATGCAGCTGCGCAAGGGCCCCAACGTGGTGGGCCCCTACGGCCTGCTGCAGCCCTTCGCCGACGCCATCAAGCTGTTCACCAAGGAGCCCCTGAAGCCCGCCACCAGCACCATCACCCTGTACATCACCGCCCCCACCCTGGCCCTGACCATCGCCCTGCTGCTGTGGACCCCCCTGCCCATGCCCAACCCCCTGGTGAACCTGAACCTGGGCCTGCTGTTCATCCTGGCCACCAGCAGCCTGGCCGTGTACAGCATCCTGTGGAGCGGCTGGGCCAGCAACAGCAACTACGCCCTGATCGGCGCCCTGCGCGCCGTGGCCCAGACCATCAGCTACGAGGTGACCCTGGCCATCATCCTGCTGAGCACCCTGCTGATGAGCGGCAGCTTCAACCTGAGCACCCTGATCACCACCCAGGAGCACCTGTGGCTGCTGCTGCCCAGCTGGCCCCTGGCCATGATGTGGTTCATCAGCACCCTGGCCGAGACCAACCGCACCCCCTTCGACCTGGCCGAGGGCGAGAGCGAGCTGGTGAGCGGCTTCAACATCGAGTACGCCGCCGGCCCCTTCGCCCTGTTCTTCATGGCCGAGTACACCAACATCATCATGATGAACACCCTGACCACCACCATCTTCCTGGGCACCACCTACGACGCCCTGAGCCCCGAGCTGTACACCACCTACTTCGTGACCAAGACCCTGCTGCTGACCAGCCTGTTCCTGTGGATCCGCACCGCCTACCCCCGCTTCCGCTACGACCAGCTGATGCACCTGCTGTGGAAGAACTTCCTGCCCCTGACCCTGGCCCTGCTGATGTGGTACGTGAGCATGCCCATCACCATCAGCAGCATCCCCCCCCAGACCTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCA 71 CPA1-ND4-3′UTRGTGCTGCCCGCCTAGAAAGGGTGAAGTGGTTGTTTCCGTGACGGACTGAGTACGGGTGCCTGTCAGGCTCTTGCGGAAGTCCATGCGCCATTGGGAGGGCCTCGGCCGCGGCTCTGTGCCCTTGCTGCTGAGGGCCACTTCCTGGGTCATTCCTGGACCGGGAGCCGGGCTGGGGCTCACACGGGGGCTCCCGCGTGGCCGTCTCGGCGCCTGCGTGACCTCCCCGCCGGCGGGATGTGGCGACTACGTCGGGCCGCTGTGGCCTGATGCTAAAACTAATCGTCCCAACAATTATGTTACTACCACTGACATGGCTTTCCAAAAAACACATGATTTGGATCAACACAACCACCCACAGCCTAATTATTAGCATCATCCCTCTACTATTTTTTAACCAAATCAACAACAACCTATTTAGCTGTTCCCCAACCTTTTCCTCCGACCCCCTAACAACCCCCCTCCTAATGCTAACTACCTGGCTCCTACCCCTCACAATCATGGCAAGCCAACGCCACTTATCCAGTGAACCACTATCACGAAAAAAACTCTACCTCTCTATGCTAATCTCCCTACAAATCTCCTTAATTATGACATTCACAGCCACAGAACTAATCATGTTTTATATCTTCTTCGAAACCACACTTATCCCCACCTTGGCTATCATCACCCGATGGGGCAACCAGCCAGAACGCCTGAACGCAGGCACATACTTCCTATTCTACACCCTAGTAGGCTCCCTTCCCCTACTCATCGCACTAATTTACACTCACAACACCCTAGGCTCACTAAACATTCTACTACTCACTCTCACTGCCCAAGAACTATCAAACTCCTGGGCCAACAACTTAATGTGGCTAGCTTACACAATGGCTTTTATGGTAAAGATGCCTCTTTACGGACTCCACTTATGGCTCCCTAAAGCCCATGTCGAAGCCCCCATCGCTGGGTCAATGGTACTTGCCGCAGTACTCTTAAAACTAGGCGGCTATGGTATGATGCGCCTCACACTCATTCTCAACCCCCTGACAAAAACACATGGCCTACCCCTTCCTTGTACTATCCCTATGGGGCATGATTATGACAAGGTCCATCTGCCTACGACAAACAGACCTAAAATCGCTCATTGCATACTCTTCAATCAGCCACATGGCCCTCGTAGTAACAGCCATTCTCATCCAAACCCCCTGGAGCTTCACCGGCGCAGTCATTCTCATGATCGCCCACGGGCTTACATCCTCATTACTATTCTGCCTAGCAAACTCAAACTACGAACGCACTCACAGTCGCATCATGATCCTCTCTCAAGGACTTCAAACTCTACTCCCACTAATGGCTTTTTGGTGGCTTCTAGCAAGCCTCGCTAACCTCGCCTTACCCCCCACTATTAACCTACTGGGAGAACTCTCTGTGCTAGTAACCACGTTCTCCTGGTCAAATATCACTCTCCTACTTACAGGACTCAACATGCTAGTCACAGCCCTATACTCCCTCTACATGTTTACCACAACACAATGGGGCTCACTCACCCACCACATTAACAACATGAAACCCTCATTCACACGAGAAAACACCCTCATGTTCATGCACCTATCCCCCATTCTCCTCCTATCCCTCAACCCCGACATCATTACCGGGTTTTCCTCTTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCAGCCCCTGTCCTCCCTTCACCCCCATTGCGTATGAGCATTTCAGAACTCCAAGGAGTCACAGGCATCTTTATAGTTCACGTTAACATATAGACACTGTTGGAAGCAGTTCCTTCTAAAAGGGTAGCCCTGGACTTAATACCAGCCGGATACCTCTGGCCCCCACCCCATTACTGTACCTCTGGAGTCACTACTGTGGGTCGCCACTCCTCTGCTACACAGCACGGCTTTTTCAAGGCTGTATTGAGAAGGGAAGTTAGGAAGAAGGGTGTGCTGGGCTAACCAGCCCACAGAGCTCACATTCCTGTCCCTTGGGTGAAAAATACATGTCCATCCTGATATCTCCTGAATTCAGAAATTAGCCTCCACATGTGCAATGGCTTTTTAAGCCAGAAGCAGGGTTCTGGGAATTTTGCAAGTTACCTGTGGCCAGGTGTGGTCTCGGTTACCAAATACGGTTACCTGCAGCTTTTTAGTCCTTTGTGCTCCCACGGGTCTACAGAGTCCCATCTGCCCAAAGGTCTTGAAGCTTGACAGGATGTTTTCGATTACTCAGTCTCCCAGGGCACTACTGGTCCGTAGGATTCGATTGGTCGGGGTAGGAGAGTTAAACAACATTTAAACAGAGTTCTCTCAAAAATGTCTAAAGGGATTGTAGGTAGATAACATCCAATCACTGTTTGCACTTATCTGAAATCTTCCCTCTTGGCTGCCCCCAGGTATTTACTGTGGAGAACATTGCATAGGAATGTCTGGAAAAAGCTTCTACAACTTGTTACAGCCTTCACATTTGTAGAAGCTTT 72CPA1-ND4-3′UTR-2*GTGCTGCCCGCCTAGAAAGGGTGAAGTGGTTGTTTCCGTGACGGACTGAGTACGGGTGCCTGTCAGGCTCTTGCGGAAGTCCATGCGCCATTGGGAGGGCCTCGGCCGCGGCTCTGTGCCCTTGCTGCTGAGGGCCACTTCCTGGGTCATTCCTGGACCGGGAGCCGGGCTGGGGCTCACACGGGGGCTCCCGCGTGGCCGTCTCGGCGCCTGCGTGACCTCCCCGCCGGCGGGATGTGGCGACTACGTCGGGCCGCTGTGGCCTGATGCTAAAACTAATCGTCCCAACAATTATGTTACTACCACTGACATGGCTTTCCAAAAAACACATGATTTGGATCAACACAACCACCCACAGCCTAATTATTAGCATCATCCCTCTACTATTTTTTAACCAAATCAACAACAACCTATTTAGCTGTTCCCCAACCTTTTCCTCCGACCCCCTAACAACCCCCCTCCTAATGCTAACTACCTGGCTCCTACCCCTCACAATCATGGCAAGCCAACGCCACTTATCCAGTGAACCACTATCACGAAAAAAACTCTACCTCTCTATGCTAATCTCCCTACAAATCTCCTTAATTATGACATTCACAGCCACAGAACTAATCATGTTTTATATCTTCTTCGAAACCACACTTATCCCCACCTTGGCTATCATCACCCGATGGGGCAACCAGCCAGAACGCCTGAACGCAGGCACATACTTCCTATTCTACACCCTAGTAGGCTCCCTTCCCCTACTCATCGCACTAATTTACACTCACAACACCCTAGGCTCACTAAACATTCTACTACTCACTCTCACTGCCCAAGAACTATCAAACTCCTGGGCCAACAACTTAATGTGGCTAGCTTACACAATGGCTTTTATGGTAAAGATGCCTCTTTACGGACTCCACTTATGGCTCCCTAAAGCCCATGTCGAAGCCCCCATCGCTGGGTCAATGGTACTTGCCGCAGTACTCTTAAAACTAGGCGGCTATGGTATGATGCGCCTCACACTCATTCTCAACCCCCTGACAAAACACATGGCCTACCCCTTCCTTGTACTATCCCTATGGGGCATGATTATGACAAGCTCCATCTGCCTACGACAAACAGACCTAAAATCGCTCATTGCATACTCTTCAATCAGCCACATGGCCCTCGTAGTAACAGCCATTCTCATCCAAACCCCCTGGAGCTTCACCGGCGCAGTCATTCTCATGATCGCCCACGGGCTTACATCCTCATTACTATTCTGCCTAGCAAACTCAAACTACGAACGCACTCACAGTCGCATCATGATCCTCTCTCAAGGACTTCAAACTCTACTCCCACTAATGGCTTTTTGGTGGCTTCTAGCAAGCCTCGCTAACCTCGCCTTACCCCCCACTATTAACCTACTGGGAGAACTCTCTGTGCTAGTAACCACGTTCTCCTGGTCAAATATCACTCTCCTACTTACAGGACTCAACATGCTAGTCACAGCCCTATACTCCCTCTACATGTTTACCACAACACAATGGGGCTCACTCACCCACCACATTAACAACATGAAACCCTCATTCACACGAGAAAACACCCTCATGTTCATGCACCTATCCCCCATTCTCCTCCTATCCCTCAACCCCGACATCATTACCGGGTTTTCCTCTTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCA 73 OPA1-opt_ND4-3′UTRGTGCTGCCCGCCTAGAAAGGGTGAAGTGGTTGTTTCCGTGACGGACTGAGTACGGGTGCCTGTCAGGCTCTTGCGGAAGTCCATGCGCCATTGGGAGGGCCTCGGCCGCGGCTCTGTGCCCTTGCTGCTGAGGGCCACTTCCTGGGTCATTCCTGGACCGGGAGCCGGGCTGGGGCTCACACGGGGGCTCCCGCGTGGCCGTCTCGGCGCCTGCGTGACCTCCCCGCCGGCGGGATGTGGCGACTACGTCGGGCCGCTGTGGCCTGATGCTGAAAGCTGATCGTGCCCACCATCATGCTGCTGCCTCTGACCTGGCTGAGCAAGAAACACATGATCTGGATCAACACCACCACGCACAGCCTGATCATCAGCATCATCCCTCTGCTGTTCTTCAACCAGATCAACAACAACCTGTTCAGCTGCAGCCCCACCTTCAGCAGCGACCCTCTGACAACACCTCTGCTGATGCTGACCACCTGGCTGCTGCCCCTCACAATCATGGCCTCTCAGAGACACCTGAGCAGCGAGCCCCTGAGCCGGAAAGAAACTGTACCTGAGCATGCTGATCTCCCTGCAGATCTCTCTGATCATGACCTTCACCGCCACCGAGCTGATCATGTTCTACATCTTTTTCGAGACAACGCTGATCCCCACACTGGCCATCATCACCAGATGGGGCAACCAGCCTGAGAGACTGAACGCCGGCACCTACTTTCTGTTCTACACCCTCGTGGGCAGCCTGCCACTGCTGATTGCCCTGATCTACACCCACAACACCCTGGGCTCCCTGAACATCCTGCTGCTGACACTGACAGCCCAAGAGCTGAGCAACAGCTGGGCCAACAATCTGATGTGGCTGGCCTACACAATGGCCTTCATGGTCAAGATGCCCCTGTACGGCCTGCACCTGTGGCTGCCTAAAGCTCATGTGGAAGCCCCTATCGCCGGCTCTATGGTGCTGGCTGCAGTGCTGCTGAAACTCGGCGGCTACGGCATGATGCGGCTGACCCTGATTCTGAATCCCCTGACCAAGCACATGGCCTATCCATTTCTGGTGCTGAGCCTGTGGGGCATGATTATGACCAGCAGCATCTGCCTGCGGCAGACCGATCTGAAGTCCCTGATCGCCTACAGCTCCATCAGCCACATGGCCCTGGTGGTCACCGCCATCCTGATTCAGACCCCTTGGAGCTTTACAGGCGCCGTGATCCTGATGATTGCCCACGGCCTGACAAGCAGCCTGCTGTTTTGTCTGGCCAACAGCAACTACGAGCGGACCCACAGCAGAATCATGATCCTGTCTCAGGGCCTGCAGACCCTCCTGCCTCTTATGGCTTTTTGGTGGCTGCTGGCCTCTCTGGCCAATCTGGCACTGCCTCCTACCATCAATCTGCTGGGCGAGCTGAGCGTGCTGGTCACCACATTCAGGTGGTCCAATATCACCCTGCTGCTCACCGGCCTGAACATGCTGGTTACAGCCCTGTACTCCCTGTACATGTTCACCACCACACAGTGGGGAAGCCTGACACACCACATCAACAATATGAAGCCCAGCTTCACCCGCGAGAACACCCTGATGTTCATGCATCTGAGCCCCATTCTGCTGCTGTCCCTGAATCCTGATATCATCACCGGCTTCTCCAGCTGAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATTGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGCCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCAGCCCCTGTCCTCCCTTCACCCCCATTGCGTATGAGCATTTCAGAACTCCAAGGAGTCACAGGCATCTTTATAGTTCACGTTAACATATAGACACTGTTGGAAGCAGTTCCTTCTAAAAGGGTAGCCCTGGACTTAATACCAGCCGGATACCTCTGGCCCCCACCCCATTACTGTACCTCTGGAGTCACTACTGTGGGTCGCCACTCCTCTGCTACACAGCACGGCTTTTTCAAGGCTGTATTGAGAAGGGAAGTTAGGAAGAAGGGTGTGCTGGGCTAACCAGCCCACAGAGCTCACATTCCTGTCCCTTGGGTGTAAAAATACATGTCCATCCTGATATCTCCTGAATTCAGAAATTAGCCTCCACATGTGCAATGGCTTTAAGAGCCAGAAGCAGGGTTCTGGGAATTTTGCAAGTTACCTGTGGCCAGGTGTGGTCTCGGTTACCAAATACGGTTACCTGCAGCTTTTTAGTCCTTTGTGCTCCCACGGGTCTACAGAGTCCCATCTGCCCAAAGGTCTTGAAGCTTGACAGGATGTTTTCGATTACTCAGTCTCCCAGGGCACTACTGGTCCGTAGGATTCGATTGGTCGGGGTAGGAGAGTTAAACAACATTTAAACAGAGTTCTCTCAAAAATGTCTAAAGGGATTGTAGGTAGATAACATCCAATCACTGTTTGCACTTATCTGAAATCTTCCCTCTTGGCTGCCCCCAGGTATTTACTGTGGAGAACATTGCATAGGAATGTCTGGAAAAAGCTTCTACAACTTGTTACAGCCTTCACATTTGTAGAAGCTTT 74 OPA1-opt_ND4-3′UTFR*GTGCTGCCCGCCTAGAAAGGGTGAAGTGGTTGTTTCCGTGACGGACTGAGTACGGGTGCCTGTCAGGCTCTTGCGGAAGTCCATGCGCCATTGGGAGGGCCTCGGCCGCGGCTCTGTGCCCTTGCTGCTGAGGGCCACTTCCTGGGTCATTCCTGGACCGGGAGCCGGGCTGGGGCTCACACGGGGGCTCCCGCGTGGCCGTCTCGGCGCCTGCGTGACCTCCCCGCCGGCGGGATGTGGCGACTACGTCGGGCCGCTGTGGCCTGATGCTGAAGCTGATCGTGCCCACCATCATGCTGCTGCCTCTGACCTGGCTGAGCAAGAAACACATGATCTGGATCAACACCACCACGCACAGCCTGATCATCAGCATCATCCCTCTGCTGTTCTTCAACCAGATCAACAACAACCTGTTCAGCTGCAGCCCCACCTTCAGCAGCGACCCTCTGACAACACCTCTGCTGATGCTGACCACCTGGCTGCTGCCCCTCACAATCATGGCCTCTCAGAGACACCTGAGCAGCGAGCCCCTGASCCGGAGAAACTSTACCTGAGCATSCTSATCTCCCTGCAGATCTCTCTGATCATSACCTTCACCCCCACCGAGCTGATCATGTTCTACATCTTTTTCGAGACAACGCTGATCCCCACACTGGCCATCATCACCAGATGGGGCAACCAGCCTGAGAGACTGAACGCCGGCACCTACTTTCTGTTCTACACCCTCGTGGGCAGCCTGCCACTGCTGATTGCCCTGATCTACACCCACAACACCCTGGGCTCCCTGAACATCCTGCTGCTGACACTGACAGCCCAAGAGCTGAGCAACAGCTGGGCCAACAATCTGATGTGGCTGGCCTACACAATGGCCTTCATGGTCAAGATGCCCCTGTACGGCCTGCACCTGTGGCTGCCTAAAGCTCATGTGGAAGCCCCTATCGCCGGCTCTATGGTGCTGGCTGCAGTGCTGCTGAAACTCGGCGGCTACGGCATGATGCGGCTGACCCTGATTCTGAATCCCCTGACCAAGCACATGGCCTATCCATTTCTGGTGCTGAGCCTGTGGGGCATGATTATGACCAGCAGCATCTGCCTGCGGCAGACCGATCTGAAGTCCCTGATCGCCTACAGCTCCATCAGCCACATGGCCCTGGTGGTCACCGCCATCCTGATTCAGACCCCTTGGAGCTTTACAGGCGCCGTGATCCTGATGATTGCCCACGGCCTGACAAGCAGCCTGCTGTTTTGTCTGGCCAACAGCAACTACGAGCGGACCCACAGCAGAATCATGATCCTGTCTCAGGGCCTGCAGACCCTCCTGCCTCTTATGGCTTTTTGGTGGCTGCTGGCCTCTCTGGCCAATCTGGCACTGCCTCCTACCATCAATCTGCTGGGCGAGCTGAGCGTGCTGGTCACCACATTCAGCTGGTCCAATATCACCCTGCTGCTCACCGGCCTGAACATGCTGGTTACAGCCCTGTACTCCCTGTACATGTTCACCACCACACAGTGGGGAAGCCTGACACACCACATCAACAATATGAAGCCCAGCTTCACCCGCGAGAAACACCCTGATGTTCATGCATCTGAGCCCCATTCTGCTGCTGTCCCTGAATCCTGATATCATCACCGGCTTCTCCAGCTGAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCTGCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCA 75 OPA1-opt_ND4*-3′UTRGTGCTGCCCGCCTAGAAAGGGTGAAGTGGTTGTTTCCGTGACGGACTGAGTACGGGTGCCTGTCAGGCTCTTGCGGAAGTCCATGCGCCATTGGGAGGGCCTCGGCCGCGGCTCTGTGCCCTTGCTGCTGAGGGCCACTTCCTGGGTCATTCCTGGACCGGGAGCCGGGCTGGGGCTCACACGGGGGCTCCCGCGTGGCCGTCTCGGCGCCTGCGTGACCTCCCCGCCGGCGGGATGTGGCGACTACGTCGGGCCGCTGTGGCCTGATGCTGAAGCTGATCGTGCCCACCATCATGCTGCTGCCCCTGACCTGGCTGAGCAAGAAGCACATGATCTGGATCAACACCACCACCCACAGCCTGATCATCAGCATCATCCCCCTGCTGTTCTTCAACCAGATCAACAACAACCTGTTTCAGCTGCAGCCCCACCTTCAGCAGCGACCCCCTGACCACCCCCCTGCTGATGCTGACCACCTGGCTGCTGCCCCTGACCATCATGGCCAGCCAGCGCCACCTGAGCAGCGAGCCCCTGAGCCGCAAGAAGCTGTACCTGAGCATGCTGATCAGCCTGCAGATCAGCCTGATCATGACCTTCACCGCCACCGAGCTGATCATGTTCTACATCTTCTTCGAGACCACCCTGATCCCCACCCTGGCCATCATCACCCGCTGGGGCAACCAGCCCGAGCGCCTGAACGCCGGCACCTACTTCCTGTTCTACACCCTGGTGGGCAGCCTGCCCCTGCTGATCGCCCTGATCTACACCCACAACACCCTGGGCAGCCTGAACATCCTGCTGCTGACCCTGACCGCCCAGGAGCTGAGCAACAGCTGGGCCAACAACCTGATGTGGCTGGCCTACACCATGGCCTTCATGGTGAAGATGCCCCTGTACGGCCTGCACCTGTGGCTGCCCAAGGCCCACGTGGAGGCCCCCATCGCCGGCAGCATGGTGCTGGCCGCCGTGCTGCTGAAGCTGGGCGGCTACGGCATGATGCGCCTGACCCTGATCCTGAACCCCCTGACCAAGCACATGGCCTACCCCTTCCTGGTGCTGAGCCTGTGGGGCATGATCATGACCAGCAGCATCTGCCTGCGCCAGACCGACCTGAAGAGCCTGATCGCCTACAGCAGCATCAGCCACATGGCCCTGGTGGTGACCGCCATCCTGATCCAGACCCCCTGGAGCTTCACCGGCGCCGTGATCCTGATGATCGCCCACGGCCTGACCAGCAGCCTGCTGTTCTGCCTGGCCAACAGCAACTACGAGCGCACCCACAGCCGCATCATGATCCTGAGCCAGGGCCTGCAGACCCTGCTGCCCCTGATGGCCTTCTGGTGGCTGCTGGCCAGCCTGGCCAACCTGGCCCTGCCCCCCACCATCAACCTGCTGGGCGAGCTGAGCGTGCTGGTGACCACCTTCAGCTGGAGCAACATCACCCTGCTGCTGACCGGCCTGAACATGCTGGTGACCGCCCTGTACAGCCTGTACATGTTCACCACCACCCAGTGGGGCAGCCTGACCCACCACATCAACAACATGAAGCCCAGCTTCACCCGCGAGAACACCCTGATGTTCATGCACCTGAGCCCCATCCTGCTGCTGAGCCTGAACCCCGACATCATCACCGGCTTCAGCAGCTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCAGCCCCTGTCCTCCCTTCACCCCCATTGCGTATGAGCATTTCAGAACTCCAAGGAGTCACAGGCATCTTTATAGTTCACGTTAACATATAGACACTGTTGGAAGCAGTTCCTTCTAAAAGGGTAGCCCTGGACTTAATACCAGCCGGATACCTCTGGCCCCCACCCCATTACTGTACCTCTGGAGTCACTACTGTGGGTCGCCACTCCTCTGCTACACAGCACGGCTTTTTCAAGGCTGTATTGAGATGGGAAAGTTAGGAAGAAGGGTGTGCTGGGCTbACCAGCCCACAGAGCTCACATTCCTGTCCCTTGGGTGAAAAATACATGTCCATCCTGATATCTCCTGAATTCAGAAATTAGCCTCCACATGTGCAATGGCTTTAAGAGCCAGAAGCAGGGTTCTGGGAATTTTGCAAGTTACCTGTGGCCAGGTGTGGTCTCGGTTACCAAATACGGTTACCTGCAGCTTTTTAGTCCTTTGTGCTCCCACGGGTCTACAGAGTCCCATCTGCCCAAAGGTCTTGAAAGCTTGACAGGATGTTTTCGATTACTCAGTCTCCCAGGGCACTACTGGTCCGTAGGATTCGATTGGTCGGGGTAGGAGAGTTAAACAACATTTAAACAGAGTTCTCTCAAAAATGTCTAAAGGGATTGTAGGTAGATAACATCCAATCACTGTTTGCACTTATCTGAAATCTTCCCTCTTGGCTGCCCCCAGGTATTTACTGTGGAGAACATTGCATAGGAATGTCTGGAAAAAGCTTCTACAACTTGTTACAGCCTTCACATTTGTAGAAGCTTT 76 OPA1-opt_ND4*-3′UTRGTGCTGCCCGCCTAGAAAGGGTGAAGTGGTTGTTTCCGTGACGGACTGAGTACGGGTGCCTGTCAGGCTCTTGCGGAAGTCCATGCGCCATTGGGAGGGCCTCGGCCGCGGCTCTGTGCCCTTGCTGCTGAGGGCCACTTCCTGGGTCATTCCTGGACCGGGAGCCGGGCTGGGGCTCACACGGGGGCTCCCGCGTGGCCGTCTCGGCGCCTGCGTGACCTCCCCGCCGGCGGGATGTGGCGACTACGTCGGGCCGCTGTGGCCTGATGCTGAAGCTGATCGTGCCCACCATCATGCTGCTGCCCCTGACCTGGCTGAGCAAGAAGCACATGATCTGGATCAACACCACCACCCACAGCCTGATCATCAGCATCATCCCCCTGCTGTTCTTCAACCAGATCAACAACAACCTGTTCAGCTGCAGCCCCACCTTCAGCAGCGACCCCCTGACCACCCCCCTGCTGATGCTGACCACCTGGCTGCTGCCCCTGACCATCATGGCCAGCCAGCGCCACCTGAGCAGCGAGCCCCTGAGCCGCAAGAAGCTGTACCTGAGCATGCTGATCAGCCTGCAGATCAGCCTGATCATGACCTTCACCGCCACCGAGCTGATCATGTTCTACATCTTCTTCGAGACCACCCTGATCCCCACCCTGGCCATCATCACCCGCTGGGGCAACCAGCCCGAGCGCCTGAACGCCGGCACCTACTTCCTGTTCTACACCCTGGTGGGCAGCCTGCCCCTGCTGATCGCCCTGATCTACACCCACAACACCCTGGGCAGCCTGAACATCCTGCTGCTGACCCTGACCGCCCAGGAGCTGAGCAACAGCTGGGCCAACAACCTGATGTGGCTGGCCTACACCATGGCCTTCATGGTGAAGATGCCCCTGTACGGCCTGCACCTGTGGCTGCCCAAGGCCCACGTGGAGGCCCCCATCGCCGGCAGCATGGTGCTGGCCGCCGTGCTGCTGAAGCTGGGCGGCTACGGCATGATGCGCCTGACCCTGATCCTGAACCCCCTGACCAAGCACATGGCCTACCCCTTCCTGGTGCTGAGCCTGTGGGGCATGATCATGACCAGCAGCATCTGCCTGCGCCAGACCGACCTGAAGAGCCTGATCGCCTACAGCAGCATCAGCCACATGGCCCTGGTGGTGACCGCCATCCTGATCCAGACCCCCTGGAGCTTCACCGGCGCCGTGATCCTGATGATCGCCCACGGCCTGACCAGCAGCCTGCTGTTCTGCCTGGCCAACAGCAACTACGAGCGCACCCACAGCCGCATCATGATCCTGAGCCAGGGCCTGCAGACCCTGCTGCCCCTGATGGCCTTCTGGTGGCTGCTGGCCAGCCTGGCCAACCTGGCCCTGCCCCCCACCATCAACCTGCTGGGCGAGCTGAGCGTGCTGGTGACCACCTTCAGCTGGAGCAACATCACCCTGCTGCTGACCGGCCTGAACATGCTGGTGACCGCCCTGTACAGCCTGTACATGTTCACCACCACCCAGTGGGGCAGCCTGACCCACCACATCAACAACATGAAGCCCAGCTTCACCCGCGAGAACACCCTGATGTTCATGCACCTGAGCCCCATCCTGCTGCTGAGCCTGAACCCCGACATCATCACCGGCTTCAGCAGCTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCA 77 OPA1-ND6-3′UTRGTGCTGCCCGCCTAGAAAGGGTGAAGTGGTTGTTTCCGTGACGGACTGAGTACGGGTGCCTGTCAGGCTCTTGCGGAAGTCCATGCGCCATTGGGAGGGCCTCGGCCGCGGCTCTGTGCCCTTGCTGCTGAGGGCCACTTCCTGGGTCATTCCTGGACCGGGAGCCGGGCTGGGGCTCACACGGGGGCTCCCGCGTGGCCGTCTCGGCGCCTGCGTGACCTCCCCGCCGGCGGGATGTGGCGACTACGTCGGGCCGCTGTGGCCTGATGATGTATGCTTTGTTTCTGTTGAGTGTGGGTTTAGTAATGGGGTTTGTGGGGTTTTCTTCTAAGCCTTCTCCTATTTATGGGGGTTTAGTATTGATTGTTAGCGGTGTGGTCGGGTGTGTTATTATTCTGAATTTTGGGGGAGGTTATATGGGTTTAATGGTTTTTTTAATTTATTTAGGGGGAATGATGGTTGTCTTTGGATATACTACAGCGATGGCTATTGAGGAGTATCCTGAGGCATGGGGGTCAGGGGTTGAGGTCTTGGTGAGTGTTTTAGTGGGGTTAGCGATGGAGGTAGGATTGGTGCTGTGGGTGAAAGAGTATGATGGGGTGGTGGTTGTGGTAAACTTTAATAGTGTAGGAAGCTGGATGATTTATGAAGGAGAGGGGTCAGGGTTGATTCGGGAGGATCCTATTGGTGCGGGGGCTTTGTATGATTATGGGCGTTGGTTAGTAGTAGTTACTGGTTGGACATTGTTTGTTGGTGTATATATTGTAATTGAGATTGCTCGGGGGAATTAGGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTCAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCAGCCCCTGTCCTCCCTTCACCCCCATTGCGTATGAGCATTTCAGAACTCCAAGGAGTCACAGGCATCTTTATAGTTCACGTTAACATATAGACACTGTTGGAAGCAGTTCCTTCTAAAAGGGTAGCCCTGGACTTAATACCAGCCGGATACCTCTGGCCCCCACCCCATTACTGTACCTCTGGAGTCACTACTGTGGGTCGCCACTCCTCTGCTACACAGCACGGCTTTTTCAAGGCTGTATTGAGAAGGGAAGTTAGGAAGAAGGGTGTGCTGGGCTAACCAGCCCACAGAGCTCACATTCCTGTCCCTTGGGTGAAAAATACATGTCCATCCTGATATCTCCTGAATTCAGAAATTAGCCTCCACATGTGCAATGGCTTTAAGAGCCAGAAGCAGGGTTCTGGGAATTTTGCAAGTTACCTGTGGCCAGGTGTGGTCTCGGTTACCAAATACGGTTACCTGCAGCTTTTTAGTCCTTTGTGCTCCCACGGGTCTACAGAGTCCCATCTGCCCAAAGGTCTTGAAGCTTGACAGGATGTTTTCGATTACTCAGTCTCCCAGGGCACTACTGGTCCGTAGGATTCGATTGGTCGGGGTAGGAGAGTTAAACAACATTTAAACAGAGTTCTCTCAAAAATGTCTAAAGGGATTGTAGGTAGATAACATCCAATCACTGTTTGCACTTATCTGAAATCTTCCCTCTTGGCTGCCCCCAGGTATTTACTGTGGAGAACATTGCATAGGAATGTCTGGAAAAAGCTTCTACAACTTGTTACAGCCTTCACATTTGTAGAAGCTTT 78 OPA1-ND6-3′UTR*GTGCTGCCCGCCTAGAAAGGGTGAAGTGGTTGTTTCCGTGACGGACTGAGTACGGGTGCCTGTCAGGCTCTTGCGGAAGTCCATGCGCCATTGGGAGGGCCTCGGCCGCGGCTCTGTGCCCTTGCTGCTGAGGGCCACTTCCTGGGTCATTCCTGGACCGGGAGCCGGGCTGGGGCTCACACGGGGGCTCCCGCGTGGCCGTCTCGGCGCCTGCGTGACCTCCCCGCCGGCGGGATGTGGCGACTACGTCGGGCCGCTGTGGCCTGATGATGTATGCTTTGTTTCTGTTGAGTGTGGGTTTAGTAATGGGGTTTGTGGGGTTTTCTTCTAAGCCTTCTCCTATTTATGGGGGTTTAGTATTGATTGTTAGCGGTGTGGTCGGGTGTGTTATTATTCTGAATTTTGGGGGAGGTTATATGGGTTTAATGGTTTTTTTAATTTATTTAGGGGGAATGATGGTTGTCTTTGGATATACTACAGCGATGGCTATTGAGGAGTATCCTGAGGCATGGGGGTCAGGGGTTGAGGTCTTGGTGAGTGTTTTAGTGGGGTTAGCGATGGAGGTAGGATTGGTGCTGTGGGTGAAAGAGTATGATGGGGTGGTGGTTGTGGTAAACTTTAATAGTGTAGGAAGCTGGATGATTTATGAAGGAGAGGGGTCAGGGTTGATTCGGGAGGATCCTATTGGTGCGGGGGCTTTGTATGATTATGGGCGTTGGTTAGTAGTAGTTACTGGTTGGACATTGTTTGTTGGTGTATATATTGTAATTGAGATTGCTCGGGGGAATTAGGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCA 79OPA1-opt_ND6-3′UTRGTGCTGCCCGCCTAGAAAGGGTGAAGTGGTTGTTTCCGTGACGGACTGAGTACGGGTGCCTGTCAGGCTCTTGCGGAAGTCCATGCGCCATTGGGAGGGCCTCGGCCGCGGCTCTGTGCCCTTGCTGCTGAGGGCCACTTCCTGGGTCATTCCTGGACCGGGAGCCGGGCTGGGGCTCACACGGGGGCTCCCGCGTGGCCGTCTCGGCGCCTGCGTGACCTCCCCGCCGGCGGGATGTGGCGACTACGTCGGGCCGCTGTGGCCTGATGATGTACGCCCTGTTCCTGCTGAGCGTGGGCCTGGTGATGGGCTTCGTGGGCTTCAGCAGCAAGCCCAGCCCCATCTACGGCGGCCTGGTGCTGATCGTGAGCGGCGTGGTGGGCTGCGTGATCATCCTGAACTTCGGCGGCGGCTACATGGGCCTGATGGTGTTCCTGATCTACCTGGGCGGCATGATGGTGGTGTTCGGCTACACCACCGCCATGGCCATCGAGGAGTACCCCGAGGCCTGGGGCAGCGGCGTGGAGGTGCTGGTGAGCGTGCTGGTGGGCCTGGCCATGGAGGTGGGCCTGGTGCTGTGGGTGAAGGAGTACGACGGCGTGGTGGTGGTGGTGAACTTCAACAGCGTGGGCAGCTGGATGATCTACGAGGGCGAGGGCAGCGGCCTGATCCGCGAGGACCCCATCGGCGCCGGCGCCCTGTACGACTACGGCCGCTGGCTGGTGGTGGTGACCGGCTGGACCCTGTTCGTGGGCGTGTACATCGTGATCGAGATCGCCCGCGGCAACTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCAGCCCCTGTCCTCCCTTCACCCCCATTGCGTATGAGCATTTCAGAACTCCAAGGAGTCACAGGCATCTTTATAGTTCACGTTAACATATAGACACTGTTGGAAGCAGTTCCTTCTAAAAGGGTAGCCCTGGACTTAATACCAGCCGGATACCTCTGGCCCCCACCCCATTACTGTACCTCTGGAGTCACTACTGTGGGTCGCCACTCCTCTGCTACACAGCACGGCTTTTTCAAGGCTGTATTGAGAAGGGAAGTTAGGAAGAAGGGTGTGCTGGGCTAACCAGCCCACAGAGCTCACATTCCTGTCCCTTGGGTGAAAAATACATGTCCATCCTGATATCTCCTGAATTCAGAAATTAGCCTCCACATGTGCAATGGCTTTAAGAGCCAGAAGCAGGGTTCTGGGAATTTTGCAAGTTACCTGTGGCCAGGTGTGGTCTCGGTTACCAAATACGGTTACCTGCAGCTTTTTAGTCCTTTGTGCTCCCACGGGTCTACAGAGTCCCATCTGCCCAAAGGTCTTGAAGCTTGACAGGATGTTTTCGATTACTCAGTCTCCCAGGGCACTACTGGTCCGTAGGATTCGATTGGTCGGGGTAGGAGAGTTAAACAACATTTAAACAGAGTTCTCTCAAAAATGTCTAAAGGGATTGTAGGTAGATAACATCCAATCACTGTTTGCACTTATCTGAAATCTTCCCTCTTGGCTGCCCCCAGGTATTTACTGTGGAGAACATTGCATAGGAATGTCTGGAAAAAGCTTCTACAACTTGTTACAGCCTTCACATTTGTAGAAGCTTT 80OPA1-opt_ND6-3′UTR′GTGCTGCCCGCCTAGAAAGGGTGAAGTGGTTGTTTCCGTGACGGACTGAGTACGGGTGCCTGTCAGGCTCTTGCGGAAGTCCATGCGCCATTGGGAGGGCCTCGGCCGCGGCTCTGTGCCCTTGCTGCTGAGGGCCACTTCCTGGGTCATTCCTGGACCGGGAGCCGGGCTGGGGCTCACACGGGGGCTCCCGCGTGGCCGTCTCGGCGCCTGCGTGACCTCCCCGCCGGCGGGATGTGGCGACTACGTCGGGCCGCTGTGGCCTGATGATGTACGCCCTGTTCCTGCTGAGCGTGGGCCTGGTGATGGGCTTCGTGGGCTTCAGCAGCAAGCCCAGCCCCATCTACGGCGGCCTGGTGCTGATCGTGAGCGGCGTGGTGGGCTGCGTGATCATCCTGAACTTCGGCGGCGGCTACATGGGCCTGATGGTGTTCCTGATCTACCTGGGCGGCATGATGGTGGTGTTCGGCTACACCACCGCCATGGCCATCGAGGAGTACCCCGAGGCCTGGGGCAGCGGCGTGGAGGTGCTGGTGAGCGTGCTGGTGGGCCTGGCCATGGAGGTGGGCCTGGTGCTGTGGGTGAAGGAGTACGACGGCGTGGTGGTGGTGGTGAACTTCAACAGCGTGGGCAGCTGGATGATCTACGAGGGCGAGGGCAGCGGCCTGATCCGCGAGGACCCCATCGGCGCCGGCGCCCTGTACGACTACGGCCGCTGGCTGGTGGTGGTGACCGGCTGGACCCTGTTCGTGGGCGTGTACATCGTGATCGAGATCGCCCGCGGCAACTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGGTGGACTGCCA 81 OPA1-ND1-3′UTRGTGCTGCCCGCCTAGAAAGGGTGAAGTGGTTGTTTCCGTGACGGACTGAGTACGGGTGCCTGTCAGGCTCTTGCGGAAGTCCATGCGCCATTGGGAGGGCCTCGGCCGCGGCTCTGTGCCCTTGCTGCTGAGGGCCACTTCCTGGGTCATTCCTGGACCGGGAGCCGGGCTGGGGCTCACACGGGGGCTCCCGCGTGGCCGTCTCGGCGCCTGCGTGACCTCCCCGCCGGCGGGATGTGGCGACTACGTCGGGCCGCTGTGGCCTGATGGCCAACCTCCTACTCCTCATTGTACCCATTCTAATCGCAATGGCATTCCTAATGCTTACCGAACGAAAAATTCTAGGCTATATGCAACTACGCAAAGGCCCCAACGTTGTAGGCCCCTACGGGCTACTACAACCCTTCGCTGACGCCATAAAACTCTTCACCAAAGAGCCCCTAAAACCCGCCACATCTACCATCACCCTCTACATCACCGCCCCGACCTTAGCTCTCACCATCGCTCTTCTACTATGGACCCCCCTCCCCATGCCCAACCCCCTGGTCAACCTCAACCTAGGCCTCCTATTTATTCTAGCCACCTCTAGCCTAGCCGTTTACTCAATCCTCTGGTCAGGGTGGGCATCAAACTCAAACTACGCCCTGATCGGCGCACTGCGAGCAGTAGCCCAAACAATCTCATATGAAGTCACCCTAGCCATCATTCTACTATCAACATTACTAATGAGTGGCTCCTTTAACCTCTCCACCCTTATCACAACACAAGAACACCTCTGGTTACTCCTGCCATCATGGCCCTTGGCCATGATGTGGTTTATCTCCACACTAGCAGAGACCAACCGAACCCCCTTCGACCTTGCCGAAGGGGAGTCCGAACTAGTCTCAGGCTTCAACATCGAATACGCCGCAGGCCCCTTCGCCCTATTCTTCATGGCCGAATACACAAACATTATTATGATGAACACCCTCACCACTACAATCTTCCTAGGAACAACATATGACGCACTCTCCCCTGAACTCTACACAACATATTTTGTCACCAAGACCCTACTTCTAACCTCCCTGTTCTTATGGATTCGAACAGCATACCCCCGATTCCGCTACGACCAACTCATGCACCTCCTATGGAAAAACTTCCTACCACTCACCCTAGCATTACTTATGTGGTATGTCTCCATGCCCATTACAATCTCCAGCATTCCCCCTCAAACCTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCAGCCCCTGTCCTCCCTTCACCCCCATTGCGTATGAGCATTTCAGAACTCCAAGGAGTCACAGGCATCTTTATAGTTCACGTTAACATATAGACACTGTTGGAAGCAGTTCCTTCTAAAAGGGTAGCCCTGGACTTAATACCAGCCGGATACCTCTGGCCCCCACCCCATTACTGTACCTCTGGAGTCACTACTGTGGGTCGCCACTCCTCTGCTACACAGCACGGCTTTTTCAAGGCTGTATTGAGAAGGGAAGTTAGGAAGAAGGGTGTGCTGGGCTAACCAGCCCACAGAGCTCACATTCCTGTCCCTTGGGTGAAAAATACATGTCCATCCTGATATCTCCTGAATTCAGAAATTAGCCTCCACATGTGCAATGGCTTTAAGAGCCAGAAGCAGGGTTCTGGGAATTTTGCAAGTTACCTGTGGCCAGGTGTGGTCTCGGTTACCAAATACGGTTACCTGCAGCTTTTTAGTCCTTTGTGCTCCCACGGGTCTACAGAGTCCCATCTGCCCAAAGGTCTTGAAGCTTGACAGGATGTTTTCGATTACTCAGTCTCCCAGGGCACTACTGGTCCGTAGGATTCGATTGGTCGGGGTAGGAGAGTTAAACAACATTTAAACAGAGTTCTCTCAAAAATGTCTAAAGGGATTGTAGGTAGATAACATCCAAATCACTGTTTGCACTTATCTGAAATCTTCCCTCTTGGCTGCCCCCAGGTATTTACTGTGGAGAACATTGCATAGGAATGTCTGGAAAAAGCTTCTACAACTTGTTACAGCCTTCACATTTGTAGAAGCTTT 82 OPA1-NDI-3′UTR*GTGCTGCCCGCCTAGAAAGGGTGAAGTGGTTGTTTCCGTGACGGACTGAGTACGGGTGCCTGTCAGGCTCTTGCGGAAGTCCATGCGCCATTGGGAGGGCCTCGGCCGCGGCTCTGTGCCCTTGCTGCTGAGGGCCACTTCCTGGGTCATTCCTGGACCGGGAGCCGGGCTGGGGCTCACACGGGGGCTCCCGCGTGGCCGTCTCGGCGCCTGCGTGACCTCCCCGCCGGCGGGATGTGGCGACTACGTCGGGCCGCTGTGGCCTGATGGCCAACCTCCTACTCCTCATTGTACCCATTCTAATCGCAATGGCATTCCTAATGCTTACCGAACGAAAAATTCTAGGCTATATGCAACTACGCAAAGGCCCCAACGTTGTAGGCCCCTACGGGCTACTACAACCCTTCGCTGACGCCATAAAACTCTTCACCAAAGAGCCCCTAAAACCCGCCACATCTACCATCACCCTCTACATCACCGCCCCGACCTTAGCTCTCACCATCGCTCTTCTACTATGGACCCCCCTCCCCATGCCCAACCCCCTGGTCAACCTCAACCTAGGCCTCCTATTTATTCTAGCCACCTCTAGCCTAGCCGTTTACTCAATCCTCTGGTCAGGGTGGGCATCAAACTCAAACTACGCCCTGATCGGCGCACTGCGAGCAGTAGCCCAAACAATCTCATATGAAGTCACCCTAGCCATCATTCTACTATCAACATTACTAATGAGTGGCTCCTTTAACCTCTCCACCCTTATCACAACACAAGAACACCTCTGGTTACTCCTGCCATCATGGCCCTTGGCCATGATGTGGTTTATCTCCACACTAGCAGAGACCAACCGAACCCCCTTCGACCTTGCCGAAGGGGAGTCCGAACTAGTCTCAGGCTTCAACATCGAATACGCCGCAGGCCCCTTCGCCCTATTCTTCATGGCCGAATACACAAACATTATTATGATGAACACCCTCACCACTACAATCTTCCTAGGAACAACATATGACGCACTCTCCCCTGAACTCTACACAACATATTTTGTCACCAAGACCCTACTTCTAACCTCCCTGTTCTTATGGATTCGAACAGCATACCCCCGATTCCGCTACGACCAACTCATGCACCTCCTATGGAAAAACTTCCTACCACTCACCCTAGCATTACTTATGTGGTATGTCTCCATGCCCATTACAATCTCCAGCATTCCCCCTCAAACCTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGGTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTCTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCA 83 OPA1-opt_ND1-3′UTRGTGCTGCCCGCCTAGAAAGGGTGAAGTGGTTGTTTCCGTGACGGACTGAGTACGGGTGCCTGTCAGGCTCTTGCGGAAGTCCATGCGCCATTGGGAGGGCCTCGGCCGCGGCTCTGTGCCCTTGCTGCTGAGGGCCACTTCCTGGGTCATTCCTGGACCGGGAGCCGGGCTGGGGCTCACACGGGGGCTCCCGCGTGGCCGTCTCGGCGCCTGCGTGACCTCCCCGCCGGCGGGATGTGGCGACTACGTCGGGCCGCTGTGGCCTGATGGCCAACCTGCTGCTGCTGATCGTGCCCATCCTGATCGCCATGGCCTTCCTGATGCTGACCGAGCGCAAGATCCTGGGCTACATGCAGCTGCGCAAGGGCCCCAACGTGGTGGGCCCCTACGGCCTGCTGCAGCCCTTCGCCGACGCCATCAAGCTGTTCACCAAGGAGCCCCTGAAGCCCGCCACCAGCACCATCACCCTGTACATCACCGCCCCCACCCTGGCCCTGACCATCGCCCTGCTGCTGTGGACCCCCCTGCCCATGCCCAACCCCCTGCTGAACCTGAACCTGGGCCTGCTGTTCATCCTGGCCACCAGCAGCCTGGCCGTGTACAGCATCCTGTGGAGCGGCTGGGCCAGCAACAGCAACTACGCCCTGATCGGCGCCCTGCGCGCCGTGGCCCAGACCATCAGCTACGAGGTGACCCTGGCCATCATCCTGCTGAGCACCCTGCTGATGAGCGGCAGCTTCAACCTGAGCACCCTGATCACCACCCAGGAGCACCTGTGGCTGCTGCTGCCCAGCTGGCCCCTGGCCATGATGTGGTTCATCAGCACCCTGGCCGAGACCAACCGCACCCCCTTCGACCTGGCCGAGGGCGAGAGCGAGCTGGTGAGCGGCTTCAACATCGAGTACGCCGCCGGCCCCTTCGCCCTGTTCTTCATGGCCGAGTACACCAACATCATCATGATGAACACCCTGACCACCACCATCTTCCTGGGCACCACCTACGACGCCCTGAGCCCCGAGCTGTACACCACCTACTTCGTGACCAAGACCCTGCTGCTGACCAGCCTGTTCCTGTGGATCCGCACCGCCTACCCCCGCTTCCGCTACGACCAGCTGATGCACCTGCTGTGGAAGAACTTCCTGCCCCTGACCCTGGCCCTGCTGATGTGGTACGTGAGCATGCCCATCACCATCAGCAGCATCCCCCCCCAGACCTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGGTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTTATACAlCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCAGCCCCTGTCCTCCCTTCACCCCCATTGCGTATGAGCATTTCAGAACTCCAAGGAGTCACAGGCATCTTTATAGTTCACGTTAACATATAGACACTGTTGGAAGCAGTTCCTTCTAAAAGGGTAGCCCTGGACTTAATACCAGCCGGATACCTCTGGCCCCCACCCCATTACTGTACCTCTGGAGTCACTACTGTGGGTCGCCACTCCTCTGCTACACAGCACGGCTTTTTCAAGGCTGTATTGAGAAGGGAAGTTAGGAAGAAGGGTGTGCTGGGCTAACCAGCCCACAGAGCTCACATTCCTGTCCCTTGGGTGAAAAATACATGTCCATCCTGATATCTCCTGAATTCAGAAATTAGCCTCCACATGTGCAATGGCTTTAAGAGCCAGAAGCAGGGTTCTGGGAATTTTGCAAGTTACCTGTGGCCAGGTGTGGTCTCGGTTACCAAATACGGTTACCTGCAGCTTTTTAGTCCTTTGTGCTCCCACGGGTCTACAGAGTCCCATCTGCCCAAAGGTCTTGAAGCTTGACAGGATGTTTTCGATTACTCAGTCTCCCAGGGCACTACTGGTCCGTAGGATTCGATTGGTCGGGGTAGGAGAGTTAAACAACATTTAAACAGAGTTCTCTCAAAAATGTCTAAAGGGATTGTAGGTAGATAACATCCAATCACTGTTTGCACTTATCTGAAATCTTCCCTCTTGGCTGCCCCCAGGTATTTACTGTGGAGAACATTGCATAGGAATGTCTGGAAAAAGCTTCTACAACTTGTTACAGCCTTCACATTTGTAGAAGCTTT 84 OPA1-opt_ND1-3′UTR*GTGCTGCCCGCCTAAAAGGGTGAAGTGGTTGTTTCCGTGACGGACTGAGTACGGGTGCCTGTCAGGCTCTTGCGGAAGTCCATGCGCCATTGGGAGGGCCTCGGCCGCGGCTCTGTGCCCTTGCTGCTGAGGGCCACTTCCTGGGTCATTCCTGGACCGGGAGCCGGGCTGGGGCTCACACGGGGGCTCCCGCGTGGCCGTCTCGGCGCCTGCGTGACCTCCCCGCCGGCGGGATGTGGCGACTACGTCGGGCCGCTGTGGCCTGATGGCCAACCTGCTGCTGCTGATCGTGCCCATCCTGATCGCCATGGCCTTCCTGATGCTGACCGAGCGCAAGATCCTGGGCTACATGCAGCTGCGCAAGGGCCCCAACGTGGTGGGCCCCTACGGCCTGCTGCAGCCCTTCGCCGACGCCATCAAGCTGTTCACCAAGGAGCCCCTGAAGCCCGCCACCAGCACCATCACCCTGTACATCACCGCCCCCACCCTGGCCCTGACCATCGCCCTGCTGCTGTGGACCCCCCTGCCCATGCCCAACCCCCTGGTGAACCTGAACCTGGGCCTGCTGTTCATCCTGGCCACCAGCAGCCTGGCCGTGTACAGCATCCTGTGGAGCGGCTGGGCCAGCAACAGCAACTACGCCCTGATCGGCGCCCTGCGCGCCGTGGCCCAGACCATCAGCTACGAGGTGACCCTGGCCATCATCCTGCTGAGCACCCTGCTGATGAGCGGCAGCTTCAACCTGAGCACCCTGATCACCACCCAGGAGCACCTGTGGCTGCTGCTGCCCAGCTGGCCCCTGGCCATGATGTGGTTTCATCAGCACCCTGGCCGAGACCAACCGCACCCCCTTCGACCTGGCCGAGGGCGAGAGCGAGCTGGTGAGCGGCTTCAACATCGAGTACGCCGCCGGCCCCTTCGCCCTGTTCTTCATGGCCGAGTACACCAACATCATCATGATGAACACCCTGACCACCACCATCTTCCTGGGCACCACCTACGACGCCCTGAGCCCCGAGCTGTACACCACCTACTTCGTGACCAAGACCCTGCTGCTGACCAGCCTGTTCCTGTGGATCCGCACCGCCTACCCCCGCTTCCGCTACGACCAGCTGATGCACCTGCTGTGGAAGAACTTCCTGCCCCTGACCCTGGCCCTGCTGATGTGGTACGTGAGCATGCCCATCACCATCAGCAGCATCCCCCCCCAGACCTAAGAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAAATTATTTTTCCCTTTGAGGGTCTTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTTGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCA 85 β-actin-S primer CGAGATCGTGCGGGACAT 86β-actin-A primer CAGGAAGGAGGGCTGGAAC 87 ND4-S primer CTGCCTACGACAAACAGAC88 ND4-A primer AGTGCGTTCGTAGTTTGAG 89 ND6-F primerATGATGTATGCTTTGTTTCTG 90 ND6-R primer CTAATTCCCCCGAGCAATCTC 91ND6-S primer AGTGTGGGTTTAGTAATG 92 NDS-A primer TGCCTCAGGATACTCCTC 93β-actin-F primer CTCCATCCTGGCCTCGCTGT 94 β-actin-R primerGCTGTCACCTTCACCGTTCC 95 ND6-F primer GGGTTTTCTTCTAAGCCTTCTCC 96ND6-R primer CCATCATACTCTTTCACCCACAG 97 opt_ND6-F primerCGCCTGCTGACCGGCTGCGT 98 opt_ND6-R CCAGGCCTCGGGGTACTCCT 99 ND1-F primerATGGCCGCATCTCCGCACACT 100 ND1-R primer TTAGGTTTGAGGGGGAATGCT 101ND1-F primer AACCTCAACCTAGGCCTCCTA 102 ND1-R primerTGGCAGGAGTAACCAGAGGTG 103 ND1-F primer AGGAGGCTCTGTCTGGTATCTT; 104ND1-R primer TTTTAGGGGCTCTTTGGTGAA 105 opt-ND1-F primerGCCGCCTGCTGACCGGCTGCGT 106 opt-ND1-R primer TGATGTACAGGGTGATGGTGCTGG 107ND4-S primer GCCAACAGCAACTACGAGC 108 ND4-A primer TGATGTTGCTCCAGCTGAAAG109 opt-ND4-S primer GCCTGACCCTGATCCTGAAC 110 opt-ND4-A primerGTGCGCTCGTAGTTGCTGTT 111 hsACO2GGGCAGTGCCTCCCCGCCCCGCCGCTGGCGTCAAGTTCAGCTCCACGTGTGCCATCAGTGGATCCGATCCGTCCAGCCATGGCTTCCTATTCCAAGATGGTGTGACCAGACATGCTTCCTGGTCCCCGCTTAGCCCACGGAGTGACTGTGGTTGTGGTGGGGGGGTTCTTAAAATAACTTTTTAGCCCCCGTCTTCCTATTTTGAGTTTGGTTCAGATCTTAAGCAGCTCCATGCAACTGTATTTATTTTTGATGACAAGACTCCCATCTAAAGTTTTTCTCCTGCCTGATCATTTCATTGGTGGCTGAAGGATTCTAGAGAACCTTTTGTTCTTGCAAGGAAAACAAGAATCCAAAACCAGTGACTGTTCTGTGA 112 hsATP513GGGGTCTTTGTCCTCTGTACTGTCTCTCTCCTTGCCCCTAACCCAAAAAGCTTCATTTTTCTGTGTAGGCTGCACAAGAGCCTTGATTGAAGATATATTCTTTCTGAACAGTATTTAAGGTTTCCAATAAAATGTACACCCCTCAG 113 hsAK2TGTTGGGTCCAAGAAGGAATTTCTTTCCATCCCTGTGAGGCAATGGGTGGGAATGATAGGACAGGCAAAGAGAAGCTTCCTCAGGCTAGCAAAAATATCATTTGATGTATTGATTAAAAAAGCACTTGCTTGATGTATCTTTGGCGTGTGTGCTACTCTCATCTGTGTGTATGTGTGTTGTGTGTGTGTGTGTGTGCATGCACATATGTGTTCACTCTGCTACTTTGTAAGTTTTAGGCTAGGTTGCTTTACCAGCTGTTTACTTCTTTTTTGTTGTTGTTTTGAGACAAGGTTTCGCTCTGCCACCCTGGCTGGAGTGCAGTGGCGTGATCTTGGCTCACGGCAACCTCTGCCTCCTGGGGCTCAAGCAATTATCCCACCTCAGCCTCCTGAGCAGCTGGGACTACAGGTGCATGCCACAACACCTGGCTGATATTTGTATTTTTTGTAGAGACAGGATTTTGCCAAGTTGCCCAGGCTGGTCTTGAACTCCTAGGCTTAAGCAATCCACCCACCTTGGCCTCCTGAAGTGCCAGGATCACAGACGTGAGCCACTACACCCAGCCCAGCTGTTTACTTCTTTAACCATACTTTTGATTTTATTTTTTGACCAAAATGAACTAACCCAGGTAATCTTCCAGGGACCGCAATTCCAGAACCTCATAGTATTTCTTCCATTTCCAGCAGCTGATTAGAAGTCCAGGATCATGTGAAGTCAGGCAGGGTCACAGTTCCTGATGGCACATTATGGACAGAGAATTCCATTTTGTTTTCTAACCCATGATGAAAACCCACGTGAGTCAGTGTGTGAACAGGGATCATTAATTTTTTCCCCCTAGGTGGAAGGAAAAAGGCACTTACTTTGCAGGTTACAGAAATTACTGGGAGAGGATATCGTCATAAAAAGAGCCAGGCCAAATTGGAATATTTTTGTGATCTGCATCATGATGCTGAAAATAGCAATTATTTGGGAATTGGGTTTGAAAACTGAATTGTTGCCAGAGAATTAAACCAGGTGAAAGGTCCTTTTGAATTCAGATTGTCTTCTGAACATCCAGGCTGATCATCTGAGAGCAGTCAAATCTACTTCCCCAAAAAGAGACCAGGGTAGGTTTATTTGCTTTTATTTTTAATGTTTGCCTGTGTTTCCAAGTGTGAACAAAACAGTGTGTGATCTATTCTTGGATTCATTTTGATCAGTATTTATTCAAACCCAGTCTCTCTCCAGGACATAAAACTGAAATCAGATATGTTCTTTTTAAGCCCAAACCCTCTCCTTTCTAGATCCAACCCTTCACCCCTAATTTTATGATGGCTATAGCCATGGACTTCCCCAAGAAAAGATCACCCAGAAATAAGACCACCTGTGACAGTTACCAGCTTTTATTCATAACCTTAGCTTCCCAACTATTGAGCATTTTCTAAGGTCCCTGCTGTCTTTTGGTCTCTGGTTTGATTTGTGGCAAACAGATGAAGTAACAGACTGCTATGAAGGACCACAAAAACGGCAGCCTCTGGAAAAACCATTAGAAAGTCAGTGGCAGATCCAGTAAATAATATCGCCAGCCTCAGCATAATCTGCTGCTGACTCGATTCAGTGGACTCTAAAGTGCCCAGCCTCCTGACCTGAGCTCTCCTGCCATCTGTGAGACTACCAGAGGTCTTATCTGCTGTCCACATGGCAACTGGGCATGAGTACCTGGCCACCTTGCTTCCCTCTTTGCCTGGTCCAAGTGAGTGTCTGCTGCCTCTGTCCTGCCTTGTTTTCCTGGCTCTAAACCAACTCCACCCACTCTTAATGGAAAACTCAGTCTGGCTTTGTGTGTTTCTGGGAAGCACATGACTTCTGGGAATGGGCAAGGAAGAGGAGTGAAACAAAAACTGTCAGCTATGTGTGCCTGGTCTGGGATCCTTCTCTGGGTGACAGTGGCATCATGAATCTTAGAATCAGCTCCCC 114 hsALDH2GAATCATGCAAGCTTCCTCCCTCAGCCATTGATGGAAAGTTCAGCAAGATCAGCAACAAAACCAAGAAAAATGATCCTTGCGTGCTGAATATCTGAAAAGAGAAATTTTTCCTACAAAATCTCTTGGGTCAAGAAAGTTCTAGAATTTGAATTGATAAACATGGTGGGTTGGCTGAGGGTAAGAGTATATGAGGAACCTTTTAAACGACAACAATACTGCTAGCTTTCAGGATGATTTTTAAAAAATAGATTCAAATGTGTTATCCTCTCTCTGAAACGCTTCCTATAACTCGAGTTTATAGGGGAAGAAAAAGCTATTGTTTACAATTATATCACCATTAAGGCAACTGCTACACCCTGCTTTGTATTCTGGGCTAAGATTCATTAAAAACTAGCTGCTCTTAACTTACA 115hsCOX10GAGCACTGGGACGCCCACCGCCCCTTTCCCTCCGCTGCCAGGCGAGCATGTTGTGGTAATTCTGGAACACAAGAAGAGAAATTGCTGGGTTTAGAACAAGATTATAAACGAATTCGGTGCTCAGTGATCACTTGACAGTTTTTTTTTTTTTTAAATATTACCCAAAATGCTCCCCAAATAAGAAATGCATCAGCTCAGTCAGTGAATACAAAAAAGGAATTATTTTTCCCTTTGAGGGTCTTTATACATCTCTCCTCCAACCCCACCCTCTATTCTGTTTCTTCCTCCTCACATGGGGGTACACATACACAGCTTCCTCTTTTGGTTCCATCCTTACCACCACACCACACGCACACTCCACATGCCCAGCAGAGTGGCACTTGGTGGCCAGAAAGTGTGAGCCTCATGATCTGCTGTCTGTAGTTCTGTGAGCTCAGGTCCCTCAAAGGCCTCGGAGCACCCCCTTCCTGGTGACTGAGCCAGGGCCTGCATTTTTGGTTTTCCCCACCCCACACATTCTCAACCATAGTCCTTCTAACAATACCAATAGCTAGGACCCGGCTGCTGTGCACTGGGACTGGGGATTCCACATGTTTGCCTTGGGAGTCTCAAGCTGGACTGCCAGCCCCTGTCCTCCCTTCACCCCCATTGCGTATGAGCATTTCAGAACTCCAAGGAGTCACAGGCATCTTTATAGTTCACGTTAACATATAGACACTGTTGGAAGCAGTTCCTTCTAAAAGGGTAGCCCTGGACTTAATACCAGCCGGATACCTCTGGCCCCCACCCCATTACTGTACCTCTGGAGTCACTACTGTGGGTCGCCACTCCTCTGCTACACAGCACGGCTTTTTCAAGGCTGTATTGAGAAGGGAAGTTAGGAAGAAGGGTGTGCTGGGCTAACCAGCCCACAGAGCTCACATTCCTGTCCCTTGGGTGAAAAATACATGTCCATCCTGATATCTCCTGAATTCAGAAATTAGCCTCCACATGTGCAATGGCTTTAAGAGCCAGAAGCAGGGTTCTGGGAATTTTGCAAGTTATCCTGTGGCCAGGTGTGGTCTCGGTTACCAAATACGGTTACCTGCAGCTTTTTAGTCCTTTGTGCTCCCACGGGTCTGCAGAGTCCCATCTGCCCAAAGGTCTTGAAGCTTGACAGGATGTTTTCATTACTCAGTCTCCCAGGGCACTGCTGGTCCGTAGGGATTCATTGGTCGGGGTGGGAGAGTTAAACAACATTTAAACAGAGTTCTCTCAAAAATGTCTAAAGGGATTGTAGGTAGATAACATCCAATCACTGTTTGCACTTATCTGAAATCTTCCCTCTTGGCTGCCCCCAGGTATTTACTGTGGAGAACATTGCATAGGAATGTCTGGAAAAAGCCTCTACAACTTGTTACAGCCTTCACATTTGTACAATTCATTGATTCTCTTTTCCTTCCACAATAAAATGGTATACAAGAAC 116 hsUQCRFS1GAGACTTGGACTCAAGTCATAGGCTTCTTTCAGTCTTTATGTCACCTCAGGAGACTTATTTGAGAGGAAGCCTTCTGTACTTGAAGTTGATTTGAAATATGTAAAGAAATTGATGATGTATTTGCAAACATTAATGTGAAATAAATTGAATTTAATGTTGAATACTTTCAGGCATTCACTTAATAAAGACACTGTTAAGCACTGTTATGCTCAGTCATACACGCGAAAGGTACAATGTCTTTTAGCTAATTCTAATTAAAAATTACAGACTGGTGTACAAGATACTTGTG 117 hsNDUFV1CCCACCACCCTGGCCTGCTGTCCTGCGTCTATCCATGTGGAATGCTGGACAATAAAGCGAGTGCTGCCCACCCTCCAGCTGCC 118 hsNDUFV2TTTATATTGAACTGTAAATATGTCACTAGAGAAATAAAATATGGACTTCCAATCTACGTAAACTTA 119hsSOD2ACCACGATCGTTATGCTGAGTATGTTAAGCTCTTTATGACTGTTTTTGTAGTGGTATAGAGTACTGCAGAATACAGTAAGCTGCTCTATTGTAGCATTTCTTGATGTTGCTTAGTCACTTATTTCATAAACAACTTAATGTTCTGAATAATTTCTTACTAAACATTTTGTTATTGGGCAAGTGATTGAAAATAGTAAATGCTTTGTGTGATTGA 120 hsCOX6cTCTTGGAATATAAAGAATTTCTTCAGGTTGAATTACCTAGAAGTTTGTCACTGACTTGTGTTCCTGAACTATGACACATGAATATGTGGGCTAAGAAATAGTTCCTCTTGATAAATAAACAATTAACAAATACTTTGGACAGTAAGTCTTTCTCAGTTCTTAATGATAATGCAGGGCACTTACTAGCATAAGAATTGGTTTGGGATTTAACTGTTTATGAAGCTAACTTGATTTCCGTGTTTTGTTAAAATTTCATTGTTCTAGCACATCTTTAACTGTGATAGTT 121 hsIRP1GAGACGTGCACTTGGTCGTGCGCCCAGGGAGGAAGCCGCACCACCAGCCAGCGCAGGCCCTGGTGGAGAGGCCTCCCTGGCTGCCTCTGGGAGGGGTGCTGCCTTGTAGATGGAGCAAGTGAGCACTGAGGGTCTGGTGCCAATCCTGTAGGCACAAAACCAGAAGTTTCTACATTCTCTATTTTTGTTAATCATCTTCTCTTTTTCCAGAATTTGGAAGCTAGAATGGTGGGAATGTCAGTAGTGCCAGAAAGAGAGAACCAAGCTTGTCTTTAAAGTTACTGATCACAGGACGTTGCTTTTTCACTGTTTCCTATTAATCTTCAGCTGAACACAAGCAAACCTTCTCAGGAGGTGTCTCCTACCCTCTTATTGTTCCTCTTACGCTCTGCTCAATGAAACCTTCCTCTTGAGGGTCATTTTCCTTTCTGTATTAATTATACCAGTGTTAAGTGACATAGATAAGAACTTTGCACACTTCAAATCAGAGCAGTGATTCTCTCTTCTCTCCCCTTTTCCTTCAGAGTGAATCATCCAGACTCCTCATGGATAGGTCGGGTGTTAAAGTTGTTTTGATTATGTACCTTTTGATAGATCCACATAAAAAGAAATGTGAAGTTTTCTTTTACTATCTTTTCATTTATCAAGCAGAGACCTTTGTTGGGAGGCGGTTTGGGAGAACACATTTCTAATTTGAATGAAATGAAATCTATTTTCAGTG 122 hsMRPS12CAGAAGAAGTGACGGCTGGGGGCACAGTGGGCTGGGCGCCCCTGCAGAACATGAACCTTCCGCTCCTGGCTGCCACAGGGTCCTCCGATGCTGGCCTTTGCGCCTCTAGAGGCAGCCACTCATGGATTCAAGTCCTGGCTCCGCCTCTTCCATCAGGACCACT 123 hsATP5J2AGAGGACACACTCTGCACCCCCCCACCCCACGACCTTGGCCCGAGCCCCTCCGTGAGGAA 124 mSOD2AGCCCTTCCGCCAGGCTGTGTGTCAGGCCCGTGGTGGGTGTTTTGTAGTAGTGTAGAGCATTGCA 125hsCXA1LCTTATGTTCTGTGCGCATTCTGGCAGGAATTCTGTCTCTTCAGAGACTCATCCTCAAAACAAGACTTGACACTGTGTCCTTGCCCCAGTCCTAGGAACTGTGGCACACAGAGATGTTCATTTTAAAAACGGATTTCATGAAACACTCTTGTACTTATGTTTATAAGAGAGCACTGGGTAGCCAAGTGATCTTCCCATTCACAGAGTTAGTAAACCTCTGTACTACATGCTG 126 MTS-COX10 MAASPHTLSSRLLTGCVGGSVWYLERRT 127MTS-COX8 MSVLTRLLLRGLTRLGSAAPVRRARIHSL 128 MTS-OPA1 MWRLRRAAVA 129hsCOX10 MAASPHTLSSRLLTGCVGGSVWYLERRT 130 scRPM2MAFKSHYSKGYHRSAAQKKTATSPFDSSYQYLRQNQGLVNSDPVLHASHLHPHPVVVANVNYNNVDDILHPHDLDSSINNTNNPLTHEELLYNQNVSLRSLKQQQSTNYVNNNNNNQHRYY 131 IcSirt5MRKRSLRCHLWSANASLSPRKDEVTSRKESENLVKGKKNKKSHLHLLTFTASKTGTDSVFDVQKSKECCKELGLLFTSLIHSIGSFPFDEEPKAAAVFLPGSLPQLTVLVLAPGSGSCPTGKSTPHLAASGRNAELLRPQNSMIVRQFTCRGTISSHLCAHLRKPRDSRNMARP 132 tbNDUS7MLRRTSFNFTGRAMISRGSPEWSHRLDLKKGKKTTMMHKLGTSKPNNALQYAQMTL 133 ncQCR2MISRSALSRGSQLALRRPAAAKTACRGFAAAAASPAASYEPTTIAG 134 hsATP5G2MPELILYVAITLSVAERLVGPGHACAEPSFRSSRCSAPLCLLCSGSSSPATAPHPLKMFACSKFVSTPSLVKSTSQLLSRPLSAVVLKRPEILTDESLSSLAVSCPLTSLVSSRSFQTSAISRDIDTA 135 hsLACTBMYRLMSAVTARAAAPGGLASSCGRRGVHCRAGLPPLGHGWVGGLGLGLGLALGVKLAGGLRGAAPAQSPAAPDPEASPLAEPPQEQSLAPWSPCTPAPPCSRCFARAIESSRDLL 136 spilv1MTVLAPLRRLHTRAAFSSYGREIALQKRPLNLNSCSAVRRYGTGFSNNLRIKKLKNAFGVVRANSTKSTSTVTTASPIKYDSSFVGKTGGEEIFHDMMLKHNVKFTVFGYPGGAILPVFDAIYRSPHFEFILPRHEQAAGHA137 gmCOX2MILCPLEAFTVQHILTISVMGLLSCFRSTVLRKCSKGSSGMSRFLYTNNFQRNLISSGGNESYYGYFNRRSYTSLYMGTGTVGGITSARIRVPNVGCEGFMCSSHLSITQRNSRLIHSTSKIVPN 138 crATP6MALQQAAPRVFGLLGRAPVALGQSGILTGSSGFKNQGFNGSLQSVENHVYAQAFSTSSQEEQAAPSIQGASGMKLPGMAGSMLLGKSRSGLRTGSMVPFAAQQAMNM 139 hsOPA1MWRLRRAAVACEVCQSLVKHSSGTKGSLPLQKLHLVSRSIYHSHHPTLKLQRPQLRTSFQQFSSLTNLPLRKLKFSPIKYGYQPRRN 140 hsSDHDMAVLWRLSAVCGALGGRALLLRTPVVRPAHTSAFLQDRPTPEWCGVQHIHLSPSHH 141 hsADCK3MAAILGDTIMVAKGLVKLTQAAVETHLQHLGIGGELIMAARALQSTAVEQIGMFLGKVQGQDKHEEYFAENFGGPEGEFHPSVPHAAGASTDFSSASAPDQSAPPSLGHAHSEGPAPAYVASGPFREAGFPGQASSPLGRANGRLFANPRDSFSAMGFQRRF 142 osP0644B0624-2MALLLRHSPKLRRAHAILGCERGTVVRHFSSSTCSSLVKEDTVSSSNLHPEYAKKIGGSDFSHDRQSGKELQNFKVSPQEASRASNFMRASKYGMPITANGVHSLFSCGQVVPSRCF 143Neurospora crassa ATP9 (ncATP9)MASTRVLASRLASQMAASAKVARPAVRVAQVSKRTIQTGSPLQTLKRTQMTSIVNATTRQAFQKRA 144hsGHITM MLAARLVCLRTLPSRVFHPAFTKASPVVKNSITKNQWLLTPSRE 145 hsNDUFAB1MASRVLSAYVSRLPAAFAPLPRVRMLAVARPLSTALCSAGTQTRLGTLQPALVLAQVPGRVTQLCRQY 146hsATP5G3MFACAKLACTPSLIRAGSRVAYRPISASVLSRPEASRTGEGSTVFNGAQNGVSQLICREFQTSAISR 147crATP6_hsADCK3MALQQAAPRVFGLLGRAPVALGQSGILTGSSGFKNQGFNGSLQSVENHVYAQAFSTSSQEEQAAPSIQGASGMKLPGMAGSMLLGKSRSGLRTGSMVPFAAQQAMNMCGMAAILGDTIMVAKGLVKLTQAAVETHLQHLGIGGELIMAARALQSTAVEQIGMFLGKVQGQDKHEEYFAENFGGPEGEFHFSVPHAAGASTDFSSASAPDQSAPPSLGHAHSEGPAPAYVASGPFREAGFPGQASSPLGRANGRLFANPRDSFSAMGFQRRFGG 148ncATP9_ncATP9MASTRVLASRLASCMAASAKVARPAVRVAQVSKRTIQTGSPLQTLKRTQMTSIVNATTRQAFQKRAMASTRVLASRLASQMAASAKVARPAVRVAQVSKRTIQTGSPLQTLKRTQMTSIVNATTRQAFQKRA 149zmLOC100282174MALLRAAVSELRRRGRGALTPLPALSSLLSSLSPRSPASTRPEPNNPHADRRHVIALRRCPPLPASAVLAPELLHARGLLPRHWSHASPLSTSSSSSRPADKAQLTWVDKWIPEAARPY 150ncATP9_zmLOC100282174_spilv1_ncATP9MASTRVLASRLASQMAASAKVARPAVRVAQVSKRTIQTGSPLQTLKRTQMTSIVNATTRQAFQKRAMALLRAAVSELRRRGRGALTPLPALSSLLSSLSPRSPASTRPEPNNPHADRRHVIALRRCPPLPASAVLAPELLHARGLLPRHWSHASPLSTSSSSSRPADKAQLTWVDKWIPEAARPYMTVLAPLRRLHTRAAFSSYGREIALQKRFLNLNSCSAVRRYGTGFSNNLRIKKLKNAFGVVRANSTKSTSTVTTASPIKYDSSFVGKTGGEIFHDMMLKHNVKHVFGYPGGAILPVFDAIYRSPHFEFILPRHEQAAGHAMASTRVLASRLASQMAASAKVARPAVRVAQVSKRTIQTGSPLQTLKRTQMTSIVNATTRQAFQKRA 151zmLOC100282174_hsADCK3_crATP6_hsATP5G3MALLRAAVSELRRRGRGALTPLPALSSLTSSLSPRSPASTRPERNNPHADRRHVIALRRCPPLPASAVLAPELLHARGLLPRHWSHASPLSTSSSSSRPADKAQLTWVDKWIPEAARPYMAAILGDTIMVAKGLVKLTQAAVETHLQHLGIGGELIMAARALQSTAVEQIGMFLGKVQGQDKHEEYFAENFGGPEGEFHFSVPHAAGASTDFSSASAPDQSAPPSLGHAHSEGPAPAYVASGPFREAGFPGQASSPLGRANGRLFANPRDSFSAMGFQRRFMALQQAAPRVFGLTGRAPVALGQSGILTGSSGFKNQGFNGSLQSVENHVYAQAFSTSSQEEQAAPSIQGASGMKLPGMAGSMLLGKSRSGLRTGSMVPFAAQQAMNMMFACAKLACTPSLIRAGSRVAYRPISASVLSRPEASRTGEGSTVFNGAQNGVSQLIQREFQTSAISR 152 zmLOC100282174_hsADCK3_hsATP5G3MALLRAAVSELRRRGRGALTPLPALSSLLSSLSPRSPASTRPEPNNPHADRRHVIALRRCPPLPASAVLAPELLHARGLLPRHWSHASPLSTSSSSSRPADKAQLTWVDKWIPEAARPYMAAILGDTIMVAKGLVKLTQAAVETHLQHLGIGGELIMAARALQSTAVEQIGMFLGKVQGQDKHEEYFAENFGGPEGEFHFSVPHAAGASTDFSSASAPDCSAPPSLGHAHSEGPAPAYVASGPFREAGFPGQASSPLGRANGRLFANPRDSFSAMGFQRRFMFACAKLACTPSLIRAGSRVAYRPISASVLSRPEASRTGEGSTVFNGAQNGVSQLIQREFQTSAISR 153ncATP9_zmLOC100282174MASTRVLASRLASQMAASAKVARPAVRVAQVSKRTIQTGSPLQTLKRTQMTSIVNATTRQAFQKRAMALLRAAVSELRRRGRGALTPLPALSSLLSSLSPRSPASTRPEPNNPHADRRHVIALRRCPPLPASAVLAPELLHARGLLPRHWSHASPLSTSSSSSRPADKAQLTWVDKWIPEAARPY 154hsADCK3_zmLOC100262174_crATP6_hsATP5G3MAAILGDTIMVAKGLVKLTQAAVETHLQFTLGIGGELIMAARALQSTAVEQIGMFLGKVQGQDKHEEYFAENFGGPEGEFHFSVPHAAGASTDFSSASAPDQSAPPSLGHAHSEGPAPAYVASGPFREAGFPGQASSPLGRANGRLFANPRDSFSAMGFQRRFMALLRAAVSELRRRGRGALTPLPALSSLLSSLSPRSPASTRPERNNPHADRRHVIALRRCPPLPASAVLAPELLHARGLLPRHWSHASPLSTSSSSSRPADKAQLTWVDKWIPEAARPYMALQQAAPRVFGLLGRARVALGQSGILTGSSGFKNQGFNGSLQSVENHVYAQAFSTSSQEEQAAPSIQGASGMKLPGMAGSMLLGKSRSGLRTGSMVPFAAQQAMNMMFACAKLACTPSLIRAGSRVAYRPISASVLSRPEASRTGEGSTVFNGAQNGVSQLIQREFQTSAISR 155crATP6_hsADCK3_zmLOC100282174_hsATP5G3MALQQAAPRVFGLLGRAPVALGQSGILTGSSGFKNQGFNGSLQSVENHVYAQAFSTSSQEEQAAPSIQGASGMKLPGMAGSMLLGKSRSGLRTGSMVPFAAQQAMNMMAAILGDTIMVAKGLVKLTQAAVETHLQHLGIGGELIMAARALQSTAVEQIGMFLGKVQGQDKHEEYFAENFGGPEGEFHFSVPHAAGASTDFSSASAPDQSAPPSLGHANSEGPAPAYVASGPFREAGFPGQASSPLGRANGRLFANPRDSFSAMGFQRRFMALLRAAVSELRRRGRGALTPLPALSSLLSSLSPRSPASTRPEPNNPHADRRHVIALRRCPPLPASAVLAPELLHARGLLPRHWSHASPLSTSSSSSRPADKAQLTWVDKWIPEAARPYMFACAKLACTPSLIRAGSRVAYRPISASVLSRPEASRTGEGSTVFNGAQNGVSQLIQREFQTSAISR 156 hsADCK3_zmLOC100282174MAAILGDTIMVAKGLVKLTQAAVETHLQHLGIGGELIMAARALQSTAVEQIGMFLGKVQGQDKHEEYFAENFGGPEGEFHFSVPHAAGASTDFSSASAPDQSAPPSLGHAHSEGPAPAYVASGPFREAGFPGQASSPLGRANGRLFANPRDSFSAMGFQRRFGGMALLRAAVSELRRPGRGALTPLPALSSLLSSLSPRSPASTRPEPNNPHADRRHVIALRRCPPLPASAVLAPELLHARGLLPRHWSHASPLSTSSSSSRPADKAQLTWVDKWIPEAARPYGG 157 hsADCK_zmLOC100282174_crATP6MAAILGDTIMVAKGLVKLTQAAVETHLQFTLGIGGELIMAARALQSTAVEQIGMFLGKVQGQDKHEEYFAENFGGPEGEFHFSVPHAAGASTDFSSASAPDQSAPPSLGHAHSEGPAPAYVASGPFREAGFPGQASSPLGRANGRLFANPRDSFSAMGFQRRFGGMALLRAAVSELRRRGRGALTPLPALSSLLSSLSPRSPASTRPEPNNPHADRRHVIALRRCPPLPASAVLAPELLHARGLLPRHWSHASPLSTSSSSSRPADKACLTWVDKWIPEAARPYGGMALQQAAPRVFGLLGRAPVALGQSGILTGSSGFKNQGFNGSLQSVENHVYAQAFSTSSQEEQAAPSIQGASGMKLPGMAGSMLLGKSRSGLRTGSMVPFAAQQAMNMGG 158ncATP9_zmLOC100282174_spilv1_GNFP_ncATP9MASTRVLASRLASQMAASAKVARPAVRVAQVSKRTIQTGSPLQTLKRTQMTSIVNATTRQAFQKRAMALLRAAVSELRRRGRGALTPLPALSSLLSSLSPRSPASTRPEPNNPHADRRHVIALRRCPPLPASAVLAPELLHARGLLPRHWSHASPLSTSSSSSRPADKAQLTWVDKWIPEAARPYMTVLARLRRLHTRAAFSSYGRETALQKRFLNLNSCSAVRRYGTGFSNNLRIKKLKNAFGVVRANSTKSTSTVTTASPIKYDSSFVGKTGGEIFHDMMLKHNVKHVFGYPGGAILPVFDAIYRSPHFEFILPRHEQAAGHAVSGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEMASTRVLASRLASQMAASAKVARPAVRVAQVSKRTIQTGSPLQTLKRTQMTSIVNARQAFQKRA 159 ncATP9_zmLOC10028217_spilv1_IcSirt5_osP0644B06.24-MASTRVLASRLASCMAASAKVARPAVRVAQVSKRTIQTGSPLQTLKRTQMTSIVNATTRQAFQKRAMALLR2_hsATP5G2_ncATP9AAVSELRRRGRGALTPLPALSSLLSSLSPRSPASTRPEPNNPHADRRHVIALRRCPPLPASAVLAPELLHARGLLPRHWSHASPLSTSSSSSRPADKAQLTWVDKWIPEAARPYMTVLAPLRRLHTRAAFSSYGREIALQKRFLNLNSCSAVRRYGTGFSNNLRIKKLKNAFGVVRANSTKSTSTVTTASPIKYDSSFVGKTGGEIFHDMMLKHNVKHVFGYPGGAILPVFDAIYRSPHFEFILPRHEQAAGHAMRKRSLRCHLWSANASLSPRKDEVTSRKESENLVKGKKNKKSHLHLLLFTASKIGTDSVFDVQKSKECCKELGLLFTSLIHSIGSFPFDEEPKAAAVFLPGSLPQLTVLVLAPGSGSCPTGKSTPHLAASGRNAELLRPQNSMIVRQFTCRGTISSHLCAHLRKPHDSRNMARPMALLLRHSPKLRRAHAILGCERGTVVRHFSSSTCSSLVKEDTVSSSNLHPEYAKKIGGSDFSHDRQSGKELQNFKVSPQEASRASNFMRASKYGMPITANGVHSLFSCGQVVPSRCFMPELILYVAITLSVAERLVGPGHACAEPSFRSSRCSAPLCLLCSGSSSPATAPHPLKMFACSKFVSTPSLVKSTSQLLSRPLSAVVLKRPEILTDESLSSLAVSCPLTSLVSSRSFQTSAISRDIDTAMASTRVLASRLASQMAASAKVARPAVRVAQVSKRTIQTGSPLQTLKRTQMTSIVNATTRQAFQKRA 160 ND4MLKLIVPTIMLLPLTWLSKKHMIWINTTTHSLIISIIPLLFFNQTNNNLFSCSPTFSSDPLTTPLLMLTTWLLPLTIMASQRHLSSEPLSRKKLYLSMLISLQISLIMTFTATELIMFYIFFETTLIPTLAIITRWGNQPERLNAGTYFLFYTLVGSLPLLIALTHNTLGSLNILLLTLTAQELSNSWANNLMWLAYTMAFMVKMPLYGLHLWPKAHVEAPIAGSMVLAAVLLKLGGYGMMRLTLILNPLTKHMAYPFLVSLWGMIMTSSICLRQTDLKSLIAYSSISHMALVVTAILIQTPWSFTGAVILMIAHGLTSSLLFCLANSNYERTHSRIMILSQGLQTLLPLMAFWWLLASLANLALPPTINLLGELSVLVTTFSWSNITLLLTGLNMLVTALYSLYMFTTTQWGSLTHHINNMKPSFTRENTLMFMHLSPILLLSLNPDIITGFSS 161 ND6MMYALFLLSVGLVMGFVGFSSKPSPIYGGLVLIVSGVVGCVIILNFGGGYMGLMVFLIYLGGMMVVFGYTTAMAIEEYPEAWGSGVEVLVSVLVGLAMEVGLVLVWKEYDGVVVVVNFNSVGSWMIYEGEGSGLIREDPIGAGALYDYGRWLVVVTGWTLFVGVYIVIEIARGN 162 ND1MANLLLLIVPILIAMAFLMLTERKILGYMQLRKGPNVVGPYGLLQPFADAIKLFTKEPLKPATSTITLYITAPTLALTIALLLWTPLPMPNPLVNLNLGLLFILATSSLAVYSILWSGWASNSNYALIGALRAVAQTISYEVTLAIILLSTLLMSGSFNLSTLITTQEHLWLLLPSWPLAMMWFISTLAETNRTPFDLAEGESELVSGFNIEYAAGPFALFFMAEYTNIIMMNTLTTTIFLGTTYDALSPELYTTYFVTKTLLLTSLFLWIRTAYPRFRYDQLMHLLWKNFLPLTLALLMWYVSMPITISSIPPQT

Adeno-Associated Virus (AAV)

Adeno-associated virus (AAV) is a small virus that infects humans andsome other primate species. The compositions disclosed herein comprisesfirstly an adeno-associated virus (AAV) genome or a derivative thereof.

An AAV genome is a polynucleotide sequence which encodes functionsneeded for production of an AAV viral particle. These functions includethose operating in the replication and packaging cycle for AAV in a hostcell, including encapsidation of the AAV genome into an AAV viralparticle. Naturally occurring AAV viruses are replication-deficient andrely on the provision of helper functions in trans for completion of areplication and packaging cycle. Accordingly, the AAV genome of thevector of the invention is typically replication-deficient.

The AAV genome can be in single-stranded form, either positive ornegative-sense, or alternatively in double-stranded form. The use of adouble-stranded form allows bypass of the DNA replication step in thetarget cell and so can accelerate transgene expression.

The AAV genome may be from any naturally derived serotype or isolate orGlade of AAV. Thus, the AAV genome may be the full genome of a naturallyoccurring AAV virus. As is known to the skilled person, AAV virusesoccurring in nature may be classified according to various biologicalsystems.

Commonly, AAV viruses are referred to in terms of their serotype. Aserotype corresponds to a variant subspecies of AAV which owing to itsprofile of expression of capsid surface antigens has a distinctivereactivity which can be used to distinguish it from other variantsubspecies. Typically, a virus having a particular AAV serotype does notefficiently cross-react with neutralising antibodies specific for anyother AAV serotype. AAV serotypes include AAV1, AAV2, AAV3, AAV4, AAV5,AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV12, AAV13, AAV14, AAV15, andAAV16, also recombinant serotypes, such as Rec2 and Rec3, recentlyidentified from primate brain.

A preferred serotype of AAV for use in the invention is AAV2. Otherserotypes of particular interest for use in the invention include AAV4,AAV5 and AAV8 which efficiently transduce tissue in the eye, such as theretinal pigmented epithelium. The serotype of AAV which is used can bean AAV serotype which is not AAV4. Reviews of AAV serotypes may be foundin Choi et al (Curr Gene Ther. 2005; 5(3); 299-310) and Wu et al(Molecular Therapy. 2006; 14(3), 316-327). The sequences of AAV genomesor of elements of AAV genomes including ITR sequences, rep or cap genesfor use in the invention may be derived from the following accessionnumbers for AAV whole genome sequences: Adeno-associated virus 1NC_002077, AF063497; Adeno-associated virus 2 NC_001401;Adeno-associated virus 3 NC_001729; Adeno-associated virus 3B NC_001863;Adeno-associated virus 4 NC_001829; Adeno-associated virus 5 Y18065,AF085716; Adeno-associated virus 6 NC_001862; Avian AAV ATCC VR-865AY186198, AY629583, NC_004828; Avian AAV strain DA-1 NC 006263,AY629583; Bovine AAV NC_005889, AY388617.

AAV viruses may also be referred to in terms of clades or clones. Thisrefers to the phylogenetic relationship of naturally derived AAVviruses, and typically to a phylogenetic group of AAV viruses which canbe traced back to a common ancestor, and includes all descendantsthereof. Additionally, AAV viruses may be referred to in terms of aspecific isolate, i.e. a genetic isolate of a specific AAV virus foundin nature. The term genetic isolate describes a population of AAVviruses which has undergone limited genetic mixing with other naturallyoccurring AAV viruses, thereby defining a recognisably distinctpopulation at a genetic level.

Examples of clades and isolates of AAV that may be used in the inventioninclude: Clade A: AAV1 NC_002077, AF063497, AAV6 NC_001862, Hu. 48AY530611, Hu 43 AY530606, Hu 44 AY530607, Hu 46 AY530609; Clade B: Hu.19 AY530584, Hu. 20 AY530586, Hu 23 AY530589, Hu22 AY530588, Hu24AY530590, Hu21 AY530587, Hu27 AY530592, Hu28 AY530593, Hu 29 AY530594,Hu63 AY530624, Hu64 AY530625, Hu13 AY530578, Hu56 AY530618, Hu57AY530619, Hu49 AY530612, Hu58 AY530620, Hu34 AY530598, Hu35 AY530599,AAV2 NC_001401. Hu45 AY530608, Hu47 AY530610, Hu5I AY530613, Hu52AY530614, Hu T41 AY695378, Hu S17 AY695376, Hu T88 AY695375, Hu T71AY695374, Hu T70 AY695373, Hu T40 AY695372, Hu T32 AY695371, Hu T17AY695370, Hu LG15 AY695377; Clade C: Hu9 AY530629, Hu10 AY530576, Hul1AY530577, Hu53 AY530615, Hu55 AY530617, Hu54 AY530616, Hu7 AY530628,Hu18 AY530583, Hu15 AY530580, Hu16 AY530581, Hu25 AY530591, Hu60AY530622, Ch5 AY243021, Hu3 AY530595, Hu1 AY530575, Hu4 AY530602 Hu2,AY530585, Hu61 AY530623; Clade D: Rh62 AY530573, Rh48 AY530561, Rh54AY530567, Rh55 AY530568, Cy2 AY243020, AAV7 AF513851, Rh35 AY243000,Rh37 AY242998, Rh36 AY242999, Cy6 AY243016, Cy4 AY243018, Cy3 AY243019,Cy5 AY243017, Rh13 AY243013; Clade E: Rh38 AY530558, Hu66 AY530626, Hu42AY530605, Hu67 AY530627, Hu40 AY530603, Hu41 AY530604, Hu37 AY530600,Rh40 AY530559, Rh2 AY243007, Bb1 AY243023, Bb2 AY243022, Rh10 AY243015,Hu17 AY530582, Hu6 AY530621, Rh25 AY530557, Pi2 AY530554, Pi1 AY530553,Pi3 AY530555, Rh57 AY530569, Rh50 AY530563, Rh49 AY530562, Hu39AY530601, Rh58 AY530570, Rh61 AY530572, Rh52 AY530565, Rh53 AY530566,Rh51 AY530564, Rh64 AY530574, Rh43 AY530560, AAV8 AF513852, Rh8AY242997, Rh1 AY530556; Clade F: Hu14 (AAV9) AY530579, Hu31 AY530596,Hu32 AY530597, Clonal Isolate AAV5 Y18065, AF085716, AAV 3 NC_001729,AAV 3B NC_001863, AAV4 NC_001829, Rh34 AY243001, Rh33 AY243002, Rh32AY243003.

The skilled person can select an appropriate serotype, Glade, clone orisolate of AAV for use in the present invention on the basis of theircommon general knowledge. For instance, the AAV5 capsid has been shownto transduce primate cone photoreceptors efficiently as evidenced by thesuccessful correction of an inherited color vision defect (Mancuso etal., Nature 2009, 461:784-7).

It should be understood however that the invention also encompasses useof an AAV genome of other serotypes that may not yet have beenidentified or characterised. The AAV serotype determines the tissuespecificity of infection (or tropism) of an AAV virus. Accordingly,preferred AAV serotypes for use in AAV viruses administered to patientsin accordance with the invention are those which have natural tropismfor or a high efficiency of infection of target cells within eye inLHON. Thus, AAV serotypes for use in AAV viruses administered topatients can be ones which infect cells of the neurosensory retina andretinal pigment epithelium.

Typically, the AAV genome of a naturally derived serotype or isolate orGlade of AAV comprises at least one inverted terminal repeat sequence(ITR). An ITR sequence acts in cis to provide a functional origin ofreplication, and allows for integration and excision of the vector fromthe genome of a cell. In preferred embodiments, one or more ITRsequences flank the polynucleotide sequence encoding ND4, ND6, or ND1 ora variant thereof. Preferred ITR sequences are those of AAV2, andvariants thereof. The AAV genome typically also comprises packaginggenes, such as rep and/or cap genes which encode packaging functions foran AAV viral particle. The rep gene encodes one or more of the proteinsRep78, Rep68, Rep52 and Rep40 or variants thereof. The cap gene encodesone or more capsid proteins such as VP1, VP2 and VP3 or variantsthereof. These proteins make up the capsid of an AAV viral particle.Capsid variants are discussed below.

A promoter will be operably linked to each of the packaging genes.Specific examples of such promoters include the p5, p19 and p40promoters (Laughlin et al., 1979, PNAS, 76:5567-5571). For example, thep5 and p19 promoters are generally used to express the rep gene, whilethe p40 promoter is generally used to express the cap gene.

As discussed above, the AAV genome used in the vector of the inventionmay therefore be the full genome of a naturally occurring AAV virus. Forexample, a vector comprising a full AAV genome may be used to prepareAAV virus in vitro. However, while such a vector may in principle beadministered to patients, this will be done rarely in practice.Preferably the AAV genome will be derivatised for the purpose ofadministration to patients. Such derivatisation is standard in the artand the present invention encompasses the use of any known derivative ofan AAV genome, and derivatives which could be generated by applyingtechniques known in the art. Derivatisation of the AAV genome and of theAAV capsid are reviewed in Coura and Nardi (Virology Journal, 2007,4:99), and in Choi et al and Wu et al, referenced above.

Derivatives of an AAV genome include any truncated or modified forms ofan AAV genome which allow for expression of a ND4, ND6, or ND1 transgenefrom a vector of the invention in vivo. Typically, it is possible totruncate the AAV genome significantly to include minimal viral sequenceyet retain the above function. This is preferred for safety reasons toreduce the risk of recombination of the vector with wild-type virus, andalso to avoid triggering a cellular immune response by the presence ofviral gene proteins in the target cell.

Typically, a derivative will include at least one inverted terminalrepeat sequence (ITR), preferably more than one ITR, such as two ITRs ormore. One or more of the ITRs may be derived from AAV genomes havingdifferent serotypes, or may be a chimeric or mutant ITR. A preferredmutant ITR is one having a deletion of a trs (terminal resolution site).This deletion allows for continued replication of the genome to generatea single-stranded genome which contains both coding and complementarysequences i.e. a self-complementary AAV genome. This allows for bypassof DNA replication in the target cell, and so enables acceleratedtransgene expression.

The one or more ITRs will preferably flank the polynucleotide sequenceencoding ND4, ND6, ND1, or a variant thereof at either end. Theinclusion of one or more ITRs is preferred to aid concatamer formationof the vector of the invention in the nucleus of a host cell, forexample following the conversion of single-stranded vector DNA intodouble-stranded DNA by the action of host cell DNA polymerases. Theformation of such episomal concatamers protects the vector constructduring the life of the host cell, thereby allowing for prolongedexpression of the transgene in vivo.

In preferred embodiments, ITR elements will be the only sequencesretained from the native AAV genome in the derivative. Thus, aderivative will preferably not include the rep and/or cap genes of thenative genome and any other sequences of the native genome. This ispreferred for the reasons described above, and also to reduce thepossibility of integration of the vector into the host cell genome.Additionally, reducing the size of the AAV genome allows for increasedflexibility in incorporating other sequence elements (such as regulatoryelements) within the vector in addition to the transgene.

With reference to the AAV2 genome, the following portions couldtherefore be removed in a derivative of the invention: One invertedterminal repeat (ITR) sequence, the replication (rep) and capsid (cap)genes (NB: the rep gene in the wildtype AAV genome should not to beconfused with ND4, ND6, or ND1, the human gene affected in LHON).However, in some embodiments, including in vitro embodiments,derivatives may additionally include one or more rep and/or cap genes orother viral sequences of an AAV genome. Naturally occurring AAV virusintegrates with a high frequency at a specific site on human chromosome19, and shows a negligible frequency of random integration, such thatretention of an integrative capacity in the vector may be tolerated in atherapeutic setting.

Where a derivative genome comprises genes encoding capsid proteins i.e.VP1. VP2 and/or VP3, the derivative may be a chimeric, shuffled orcapsid-modified derivative of one or more naturally occurring AAVviruses. In particular, the invention encompasses the provision ofcapsid protein sequences from different serotypes, clades, clones, orisolates of AAV within the same vector i.e. pseudotyping.

Chimeric, shuffled or capsid-modified derivatives will be typicallyselected to provide one or more desired functionalities for the viralvector. Thus, these derivatives may display increased efficiency of genedelivery, decreased immunogenicity (humoral or cellular), an alteredtropism range and/or improved targeting of a particular cell typecompared to an AAV viral vector comprising a naturally occurring AAVgenome, such as that of AAV2. Increased efficiency of gene delivery maybe effected by improved receptor or co-receptor binding at the cellsurface, improved internalisation, improved trafficking within the celland into the nucleus, improved uncoating of the viral particle andimproved conversion of a single-stranded genome to double-stranded form.Increased efficiency may also relate to an altered tropism range ortargeting of a specific cell population, such that the vector dose isnot diluted by administration to tissues where it is not needed.

Chimeric capsid proteins include those generated by recombinationbetween two or more capsid coding sequences of naturally occurring AAVserotypes. This may be performed for example by a marker rescue approachin which non-infectious capsid sequences of one serotype arecotransfected with capsid sequences of a different serotype, anddirected selection is used to select for capsid sequences having desiredproperties. The capsid sequences of the different serotypes can bealtered by homologous recombination within the cell to produce novelchimeric capsid proteins.

Chimeric capsid proteins also include those generated by engineering ofcapsid protein sequences to transfer specific capsid protein domains,surface loops or specific amino acid residues between two or more capsidproteins, for example between two or more capsid proteins of differentserotypes.

Shuffled or chimeric capsid proteins may also be generated by DNAshuffling or by error-prone PCR. Hybrid AAV capsid genes can be createdby randomly fragmenting the sequences of related AAV genes e.g. thoseencoding capsid proteins of multiple different serotypes and thensubsequently reassembling the fragments in a self-priming polymerasereaction, which may also cause crossovers in regions of sequencehomology. A library of hybrid AAV genes created in this way by shufflingthe capsid genes of several serotypes can be screened to identify viralclones having a desired functionality. Similarly, error prone PCR may beused to randomly mutate AAV capsid genes to create a diverse library ofvariants which may then be selected for a desired property.

The sequences of the capsid genes may also be genetically modified tointroduce specific deletions, substitutions or insertions with respectto the native wild-type sequence. In particular, capsid genes may bemodified by the insertion of a sequence of an unrelated protein orpeptide within an open reading frame of a capsid coding sequence, or atthe N- and/or C-terminus of a capsid coding sequence.

The unrelated protein or peptide may advantageously be one which acts asa ligand for a particular cell type, thereby conferring improved bindingto a target cell or improving the specificity of targeting of the vectorto a particular cell population. An example might include the use of RGDpeptide to block uptake in the retinal pigment epithelium and therebyenhance transduction of surrounding retinal tissues (Cronin et al., 2008ARVO Abstract: D1048). The unrelated protein may also be one whichassists purification of the viral particle as part of the productionprocess i.e. an epitope or affinity tag. The site of insertion willtypically be selected so as not to interfere with other functions of theviral particle e.g. internalisation, trafficking of the viral particle.The skilled person can identify suitable sites for insertion based ontheir common general knowledge. Particular sites are disclosed in Choiet al, referenced above.

The invention additionally encompasses the provision of sequences of anAAV genome in a different order and configuration to that of a nativeAAV genome. The invention also encompasses the replacement of one ormore AAV sequences or genes with sequences from another virus or withchimeric genes composed of sequences from more than one virus. Suchchimeric genes may be composed of sequences from two or more relatedviral proteins of different viral species.

The vector of the invention takes the form of a polynucleotide sequencecomprising an AAV genome or derivative thereof and a sequence encodingND4, ND6, ND1 or a variant thereof.

For the avoidance of doubt, the invention also provides an AAV viralparticle comprising a vector of the invention. The AAV particles of theinvention include transcapsidated forms wherein an AAV genome orderivative having an ITR of one serotype is packaged in the capsid of adifferent serotype. The AAV particles of the invention also includemosaic forms wherein a mixture of unmodified capsid proteins from two ormore different serotypes makes up the viral envelope The AAV particlealso includes chemically modified forms bearing ligands adsorbed to thecapsid surface. For example, such ligands may include antibodies fortargeting a particular cell surface receptor.

The invention additionally provides a host cell comprising a vector orAAV viral particle of the invention.

Recombinant Nucleic Acid Sequences

Also disclosed herein are recombinant nucleic acid sequences comprisinga polynucleotide sequence encoding a NADH dehydrogenase subunit-4 (ND4),NADH dehydrogenase subunit-1 (ND1) and NADH dehydrogenase subunit-6(ND6) polypeptide or a variant thereof.

The polynucleotide sequence for ND4 is shown in SEQ ID NO: 6 and encodesthe protein shown in SEQ ID NO: 160. Further nucleic acid sequences forND4 are SEQ ID NO: 7 and 8. The polynucleotide sequence for ND6 is shownin SEQ ID NO: 9 and encodes the protein shown in SEQ ID NO: 161. Afurther nucleic acid sequence for ND6 is SEQ ID NO: 10. Thepolynucleotide sequence for ND1 is shown in SEQ ID NO: 11 and encodesthe protein shown in SEQ ID NO: 162. A further nucleic acid sequence forND1 is SEQ ID NO: 12.

A variant of any one of SEQ ID NO: 160, 161, or 162 may comprisetruncations, mutants or homologues thereof, and any transcript variantsthereof which encode a functional ND4, ND6, or ND1 polypeptide. Anyhomologues mentioned herein are typically at least 70% homologous to arelevant region of ND4, ND6, or ND1, and can functionally compensate forthe polypeptide deficiency.

Homology can be measured using known methods. For example the UWGCGPackage provides the BESTFIT program which can be used to calculatehomology (for example used on its default settings) (Devereux et at(1984) Nucleic Acids Research 12, 387-395). The PILEUP and BLASTalgorithms can be used to calculate homology or line up sequences(typically on their default settings), for example as described inAltschul S. F. (1993) J Mol Evol 36:290-300; Altschul, S, F et at (1990)J Mol Biol 215:403-10. Software for performing BLAST analyses ispublicly available through the National Center for BiotechnologyInformation (http://www.ncbi.nlm.nih.gov/).

In preferred embodiments, a recombinant nucleic acid sequence may encodea polypeptide which is at least 55%, 65%, 70%, 75%, 80%, 85%, 90% andmore preferably at least 95%, 97%, 99%, 99.5%, or 100% homologous to arelevant region of ND4, ND6, or ND1 (SEQ ID NO: 160, 161, or 162) overat least 20, preferably at least 30, for instance at least 40, 60, 100,200, 300, 400 or more contiguous amino acids, or even over the entiresequence of the recombinant nucleic acid. The relevant region will beone which provides for functional activity of ND4, ND6, or ND1.

Alternatively, and preferably the recombinant nucleic acid sequence mayencode a polypeptide having at least 70%, 75%, 80%, 85%, 90% and morepreferably at least 95%, 97%, 99%, 99.5%, or 100% homologous tofull-length ND4, ND6, or ND1 (SEQ ID NO: 160, 161, or 162) over itsentire sequence. Typically the recombinant nucleic acid sequence differsfrom the relevant region of ND4, ND6, or ND1 (SEQ ID NO: 160, 161, or162) by at least, or less than, 2, 5, 10, 20, 40, 50 or 60 mutations(each of which can be substitutions, insertions or deletions).

A recombinant nucleic acid ND4, ND6, or ND1 polypeptide may have apercentage identity with a particular region of SEQ ID NO: 160, 161, or162 which is the same as any of the specific percentage homology values(i.e. it may have at least 70%, 80% or 90% and more preferably at least95%, 97/o, 99% identity) across any of the lengths of sequence mentionedabove.

Variants of ND4, ND6, or ND1 (SEQ ID NO: 160, 161, or 162) also includetruncations. Any truncation may be used so long as the variant is stillfunctional. Truncations will typically be made to remove sequences thatare non-essential for the protein activity and/or do not affectconformation of the folded protein, in particular folding of the activesite. Appropriate truncations can routinely be identified by systematictruncation of sequences of varying length from the N- or C-terminus.Preferred truncations are N-terminal and may remove all other sequencesexcept for the catalytic domain.

Variants of ND4, ND6, or ND1 (SEQ ID NO: 160, 161, or 162) furtherinclude mutants which have one or more, for example, 2, 3, 4, 5 to 10,10 to 20, 20 to 40 or more, amino acid insertions, substitutions ordeletions with respect to a particular region of ND4, ND6, or ND1 (SEQID NO: 160, 161, or 162). Deletions and insertions are made preferablyoutside of the catalytic domain as described below. Substitutions arealso typically made in regions that are non-essential for proteaseactivity and/or do not affect conformation of the folded protein.

Substitutions preferably introduce one or more conservative changes,which replace amino acids with other amino acids of similar chemicalstructure, similar chemical properties or similar side-chain volume. Theamino acids introduced may have similar polarity, hydrophilicity,hydrophobicity, basicity, acidity, neutrality or charge to the aminoacids they replace. Alternatively, the conservative change may introduceanother amino acid that is aromatic or aliphatic in the place of apre-existing aromatic or aliphatic amino acid. Conservative amino acidchanges are well known in the art and may be selected in accordance withthe properties of the amino acids.

Similarly, preferred variants of the polynucleotide sequence of ND4.ND6, or ND1 (SEQ ID NO: 6, 9, or 11) include polynucleotides having atleast 70%, 75%, 80%, 85%, 90% and more preferably at least 95%, 96%,97%, 98%, 99%, or 99.5% homologous to a relevant region of ND4, ND6, orND1 (SEQ ID NO: 6, 9, or 11). Preferably the variant displays theselevels of homology to full-length ND4, ND6, or ND1 (SEQ ID NO: 6, 9, or11) over its entire sequence.

Mitochondrial targeting sequences (MTSs) and three prime untranslatedregions (3′UTRs) can be used to target proteins or mRNA to themitochondria. The charge, length, and structure of the MTS can beimportant for protein import into the mitochondria. Particular 3′UTRsmay drive mRNA localization to the mitochondrial surface and thusfacilitate cotranslational protein import into the mitochondria.

The polynucleotide sequence for a mitochondrial targeting sequence canencode a polypeptide selected from hsCOX10, hsCOX8, scRPM2, lcSirt5,tbNDUS7, ncQCR2, hsATP5G2, hsLACTB, spilv1, gmCOX2, crATP6, hsOPA1,hsSDHD, hsADCK3, osP0644B06.24-2, Neurospora crassa ATP9 (ncATP9),hsGHITM, hsNDUFAB1, hsATP5G3, crATP6_hsADCK3, ncATP9_ncATP9,zmLOC100282174, ncATP9_zmLOC100282174_spilv1_ncATP9,zmLOCI00282174_hsADCK3_crATP6_hsATP5G3, zmLOC100282174_hsADCK3_hsATP5G3,ncATP9_zmLOC100282174, hsADCK3_zmLOC100282174_crATP6_hsATP5G3,crATP6_hsADCK3_zmLOC100282174_hsATP5G3, hsADCK3_zmLOC100282174,hsADCK3_zmLOC100282174_crATP6, ncATP9_zmLOC100282174_spily_GNFP_ncATP9,and ncATP9_zmLOC100282174_spilv1_lcSirt5_osP0644B06.24-2_hsATP5G2_ncATP9(see Table 1 for SEQ ID NO). In one example, the polynucleotidesequences, COX10 (SEQ ID NO: 1, 2, or 3) can encode the mitochondrialtargeting sequence, MTS-COX10 (SEQ ID NO: 126). In another example, thepolynucleotide sequences, COX8 (SEQ ID NO: 4) can encode themitochondrial targeting sequence, MTS-COX8 (SEQ ID NO: 127). In anotherexample, the polynucleotide sequences, OPA1 (SEQ ID NO: 5) can encodethe mitochondrial targeting sequence, MTS-OPA 1 (SEQ ID NO: 128).

The 3′UTR nucleic acid sequence can be selected from hsACO2 (SEQ ID NO:111), hsATP5B (SEQ ID NO: 112), hsAK2 (SEQ ID NO: 113), hsALDH2 (SEQ IDNO: 114), hsCOXI0 (SEQ ID NO: 115), hsUQCRFS1 (SEQ ID NO: 116), hsNDUFV1(SEQ ID NO: 117), hsNDUFV2 (SEQ ID NO: 118), hsSOD2 (SEQ ID NO: 119),hsCOX6c (SEQ ID NO: 120), hsIRPl (SEQ ID NO: 121), hsMRPS12 (SEQ ID NO:122), hsATP5J2 (SEQ ID NO: 123), mSOD2 (SEQ ID NO: 124), and hsOXA1L(SEQ ID NO: 125). The 3′UTR nucleic acid sequence can also b^(e) avariant having at least 70%, 75%, 80%, 85%, 90% and more preferably atleast 95%, 96%, 97%, 98%, 99%, 99.5%, or 100% homologous to any 3′UTRnucleic acid sequence listed here. For example, the 3′UTR nucleic acidsequence can be SEQ ID NO: 13 or 14.

Also disclosed herein are recombinant nucleic acid sequences comprisinga mitochondrial targeting sequence, a mitochondrial protein codingsequence, and a 3′UTR nucleic acid sequence. For example, therecombinant nucleic acid sequence can be selected from SEQ ID NO: 15-84.The recombinant nucleic acid sequence can also be a variant having atleast 70%, 75%, 80%, 85%, 90% and more preferably at least 95%, 96%,97%, 98%, 99%, 99.5%, or 100% homologous to any recombinant nucleic acidsequence listed here.

Promoters and Regulatory Sequences

The vector of the invention also includes elements allowing for theexpression of the disclosed transgene in vitro or in vivo. Thus, thevector typically comprises a promoter sequence operably linked to thepolynucleotide sequence encoding the ND4, ND6, or ND1 transgene or avariant thereof.

Any suitable promoter may be used. The promoter sequence may beconstitutively active i.e. operational in any host cell background, oralternatively may be active only in a specific host cell environment,thus allowing for targeted expression of the transgene in a particularcell type. The promoter may show inducible expression in response topresence of another factor, for example a factor present in a host cell.In any event, where the vector is administered for therapy, the promotermust be functional in a retinal cell background.

In some embodiments, it is preferred that the promoter showsretinal-cell specific expression in order to allow for the transgene toonly be expressed in retinal cell populations. Thus, expression from thepromoter may be retinal-cell specific, for example confined only tocells of the neurosensory retina and retinal pigment epithelium.

Preferred promoters for the ND4, ND6, or ND1 transgene include thechicken beta-actin (CBA) promoter, optionally in combination with acytomegalovirus (CME) enhancer element. In some cases, the preferredpromoters for the ND4, ND6, or ND1 transgene comprises the CAG promoter.A particularly preferred promoter is a hybrid CBA/CAG promoter, forexample the promoter used in the rAVE expression cassette. Examples ofpromoters based on human sequences that would induce retina specificgene expression include rhodospin kinase for rods and cones (Allocca etal., 2007, J Viol 81:11372-80), PR2.1 for cones only (Mancuso et al.2009. Nature) and/or RPE65 for the retinal pigment epithelium(Bainbridge et al., 2008, N Eng J Med).

The vector of the invention may also comprise one or more additionalregulatory sequences with may act pre- or post-transcriptionally. Theregulatory sequence may be part of the native ND4, ND6, or ND1 genelocus or may be a heterologous regulatory sequence. The vector of theinvention may comprise portions of the 5′UTR or 3′UTR from the nativeND4, ND6, or ND1 transcript.

Regulatory sequences are any sequences which facilitate expression ofthe transgene i.e. act to increase expression of a transcript, improvenuclear export of mRNA or enhance its stability. Such regulatorysequences include for example enhancer elements, postregulatory elementsand polyadenylation sites. A preferred polyadenylation site is theBovine Growth Hormone poly-A signal. In the context of the vector of theinvention such regulatory sequences will be cis-acting. However, theinvention also encompasses the use of trans-acting regulatory sequenceslocated on additional genetic constructs.

A preferred postregulatory element for use in a vector of the inventionis the woodchuck hepatitis postregulatory element (WPRE) or a variantthereof. Another regulatory sequence which may be used in a vector ofthe present invention is a scaffold-attachment region (SAR). Additionalregulatory sequences may be selected by the skilled person on the basisof their common general knowledge.

Preparation of Vector

The vector of the invention may be prepared by standard means known inthe art for provision of vectors for gene therapy. Thus, wellestablished public domain transfection, packaging and purificationmethods can be used to prepare a suitable vector preparation.

As discussed above, a vector of the invention may comprise the fullgenome of a naturally occurring AAV virus in addition to apolynucleotide sequence encoding ND4, ND6, or ND1 or a variant thereof.However, commonly a derivatised genome will be used, for instance aderivative which has at least one inverted terminal repeat sequence(ITR), but which may lack any AAV genes such as rcp or cap.

In such embodiments, in order to provide for assembly of the derivatisedgenome into an AAV viral particle, additional genetic constructsproviding AAV and/or helper virus functions will be provided in a hostcell in combination with the derivatised genome. These additionalconstructs will typically contain genes encoding structural AAV capsidproteins i.e. cap, VP1. VP2, VP3, and genes encoding other functionsrequired for the AAV life cycle, such as rep. The selection ofstructural capsid proteins provided on the additional construct willdetermine the serotype of the packaged viral vector.

A particularly preferred packaged viral vector for use in the inventioncomprises a derivatised genome of AAV2 in combination with AAV5 or AAV8capsid proteins. This packaged viral vector typically comprises one ormore AAV2 ITRs.

As mentioned above, AAV viruses are replication incompetent and sohelper virus functions, preferably adenovirus helper functions willtypically also be provided on one or more additional constructs to allowfor AAV replication.

All of the above additional constructs may be provided as plasmids orother episomal elements in the host cell, or alternatively one or moreconstructs may be integrated into the genome of the host cell.

In these aspects, the invention provides a method for production of avector of the invention. The method comprises providing a vector whichcomprises an adeno-associated virus (AAV) genome or a derivative thereofand a polynucleotide sequence encoding ND4, ND6, or ND1 or a variantthereof in a host cell, and providing means for replication and assemblyof the vector into an AAV viral particle. Preferably, the methodcomprises providing a vector comprising a derivative of an AAV genomeand a polynucleotide sequence encoding ND4, ND6, or ND1 or a variantthereof, together with one or more additional genetic constructsencoding AAV and/or helper virus functions. Typically, the derivative ofan AAV genome comprises at least one ITR. Optionally, the method furthercomprises a step of purifying the assembled viral particles.Additionally, the method may comprise a step of formulating the viralparticles for therapeutic use.

Methods of Therapy and Medical Uses

As discussed above, the present inventors have surprisingly demonstratedthat a vector of the invention may be used to address the cellulardysfunction underlying LHON. In particular, they have shown that use ofthe vector can correct the defect associated with LHON. This provides ameans whereby the degenerative process of the disease can be treated,arrested, palliated or prevented.

The invention therefore provides a method of treating or preventing LHONin a patient in need thereof, comprising administering a therapeuticallyeffective amount of a vector of the invention to the patient by directretinal, subretinal or intravitreal injection. Accordingly, LHON isthereby treated or prevented in the patient.

In a related aspect, the invention provides for use of a vector of theinvention in a method of treating or preventing LHON by administeringsaid vector to a patient by direct retinal, subretinal or intravitrealinjection. Additionally, the invention provides the use of a vector ofthe invention in the manufacture of a medicament for treating orpreventing LHON by direct retinal, subretinal or intravitreal injection.

In all these embodiments, the vector of the invention may beadministered in order to prevent the onset of one or more symptoms ofLHON. The patient may be asymptomatic. The subject may have apredisposition to the disease. The method or use may comprise a step ofidentifying whether or not a subject is at risk of developing, or has,LHON. A prophylactically effective amount of the vector is administeredto such a subject. A prophylactically effective amount is an amountwhich prevents the onset of one or more symptoms of the disease.

Alternatively, the vector may be administered once the symptoms of thedisease have appeared in a subject i.e. to cure existing symptoms of thedisease. A therapeutically effective amount of the antagonist isadministered to such a subject. A therapeutically effective amount is anamount which is effective to ameliorate one or more symptoms of thedisease. Such an amount may also arrest, slow or reverse some loss ofperipheral vision associated with LHON. Such an amount may also arrest,slow or reverse onset of LHON.

A typical single dose is between 10¹⁰ and 10¹² genome particles,depending on the amount of remaining retinal tissue that requirestransduction. A genome particle is defined herein as an AAV capsid thatcontains a single stranded DNA molecule that can be quantified with asequence specific method (such as real-time PCR). That dose may beprovided as a single dose, but may be repeated for the fellow eye or incases where vector may not have targeted the correct region of retinafor whatever reason (such as surgical complication). The treatment ispreferably a single permanent treatment for each eye, but repeatinjections, for example in future years and/or with different AAVserotypes may be considered.

The invention also provides a method of monitoring treatment orprevention of LHON in a patient comprising measuring activity ex vivo inretinal cells obtained from said patient following administration of theAAV vector of the invention by direct retinal, subretinal orintravitreal injection. This method can allow for determination of theefficacy of treatment.

Pharmaceutical Compositions

The vector of the invention can be formulated into pharmaceuticalcompositions. These compositions may comprise, in addition to thevector, a pharmaceutically acceptable excipient, carrier, buffer,stabiliser or other materials well known to those skilled in the art.Such materials should be non-toxic and should not interfere with theefficacy of the active ingredient. The precise nature of the carrier orother material may be determined by the skilled person according to theroute of administration, i.e. here direct retinal, subretinal orintravitreal injection.

The pharmaceutical composition is typically in liquid form. Liquidpharmaceutical compositions generally include a liquid carrier such aswater, petroleum, animal or vegetable oils, mineral oil or syntheticoil. Physiological saline solution, magnesium chloride, dextrose orother saccharide solution or glycols such as ethylene glycol, propyleneglycol or polyethylene glycol may be included. In some cases, asurfactant, such as pluronic acid (PF68) 0.001% may be used.

For injection at the site of affliction, the active ingredient will bein the form of an aqueous solution which is pyrogen-free and hassuitable pH, isotonicity and stability. Those of relevant skill in theart are well able to prepare suitable solutions using, for example,isotonic vehicles such as Sodium Chloride Injection, Ringer's Injection,Lactated Ringer's Injection. Preservatives, stabilisers, buffers,antioxidants and/or other additives may be included, as required.

For delayed release, the vector may be included in a pharmaceuticalcomposition which is formulated for slow release, such as inmicrocapsules formed from biocompatible polymers or in liposomal carriersystems according to methods known in the art.

Samples

Samples that are suitable for use in the methods described herein can benucleic acid samples from a subject. A “nucleic acid sample” as usedherein can include RNA or DNA, or a combination thereof. In anotherembodiment, a “polypeptide sample” (e.g., peptides or proteins, orfragments therefrom) can be used to ascertain information that an aminoacid change has occurred, which is the result of a genetic variant.Nucleic acids and polypeptides can be extracted from one or more samplesincluding but not limited to, blood, saliva, urine, mucosal scrapings ofthe lining of the mouth, expectorant, serum, tears, skin, tissue, orhair. A nucleic acid sample can be assayed for nucleic acid information.“Nucleic acid information,” as used herein, includes a nucleic acidsequence itself, the presence/absence of genetic variation in thenucleic acid sequence, a physical property which varies depending on thenucleic acid sequence (e.g., Tm), and the amount of the nucleic acid(e.g., number of mRNA copies). A “nucleic acid” means any one of DNA,RNA, DNA including artificial nucleotides, or RNA including artificialnucleotides. As used herein, a “purified nucleic acid” includes cDNAs,fragments of genomic nucleic acids, nucleic acids produced using thepolymerase chain reaction (PCR), nucleic acids formed by restrictionenzyme treatment of genomic nucleic acids, recombinant nucleic acids,and chemically synthesized nucleic acid molecules. A “recombinant”nucleic acid molecule includes a nucleic acid molecule made by anartificial combination of two otherwise separated segments of sequence,e.g., by chemical synthesis or by the manipulation of isolated segmentsof nucleic acids by genetic engineering techniques. As used herein, a“polypeptide” includes proteins, fragments of proteins, and peptides,whether isolated from natural sources, produced by recombinanttechniques, or chemically synthesized. A polypeptide may have one ormore modifications, such as a post-translational modification (e.g.,glycosylation, phosphorylation, etc.) or any other modification (e.g.,pegylation, etc.). The polypeptide may contain one or morenon-naturally-occurring amino acids (e.g., such as an amino acid with aside chain modification).

In some embodiments, the nucleic acid sample can comprise cells ortissue, for example, cell lines. Exemplary cell types from which nucleicacids can be obtained using the methods described herein include, butare not limited to, the following: a blood cell such as a B lymphocyte,T lymphocyte, leukocyte, erythrocyte, macrophage, or neutrophil; amuscle cell such as a skeletal cell, smooth muscle cell or cardiacmuscle cell; a germ cell, such as a sperm or egg; an epithelial cell; aconnective tissue cell, such as an adipocyte, chondrocyte; fibroblast orosteoblast; a neuron; an astrocyte; a stromal cell; an organ specificcell, such as a kidney cell, pancreatic cell, liver cell, or akeratinocyte; a stem cell; or any cell that develops therefrom. A cellfrom which nucleic acids can be obtained can be a blood cell or aparticular type of blood cell including, for example, a hematopoieticstem cell or a cell that arises from a hematopoietic stem cell such as ared blood cell, B lymphocyte, T lymphocyte, natural killer cell,neutrophil, basophil, eosinophil, monocyte, macrophage, or platelet.Generally, any type of stem cell can be used including, withoutlimitation, an embryonic stem cell, adult stem cell, or pluripotent stemcell.

In some embodiments, a nucleic acid sample can be processed for RNA orDNA isolation, for example, RNA or DNA in a cell or tissue sample can beseparated from other components of the nucleic acid sample. Cells can beharvested from a nucleic acid sample using standard techniques, forexample, by centrifuging a cell sample and resuspending the pelletedcells, for example, in a buffered solution, for example,phosphate-buffered saline (PBS). In some embodiments, after centrifugingthe cell suspension to obtain a cell pellet, the cells can be lysed toextract DNA. In some embodiments, the nucleic acid sample can beconcentrated and/or purified to isolate DNA. All nucleic acid samplesobtained from a subject, including those subjected to any sort offurther processing, are considered to be obtained from the subject. Insome embodiments, standard techniques and kits known in the art can beused to extract RNA or DNA from a nucleic acid sample, including, forexample, phenol extraction, a QIAAMP® Tissue Kit (Qiagen, Chatsworth,Calif.), a WIZARD® Genomic DNA purification kit (Promega), or a QiagenAutopure method using Puregene chemistry, which can enable purificationof highly stable DNA well-suited for archiving.

In some embodiments, determining the identity of an allele ordetermining copy number can, but need not, include obtaining a nucleicacid sample comprising RNA and/or DNA from a subject, and/or assessingthe identity, copy number, presence or absence of one or more geneticvariations and their chromosomal locations within the genomic DNA (i.e.subject's genome) derived from the nucleic acid sample.

The individual or organization that performs the determination need notactually carry out the physical analysis of a nucleic acid sample from asubject. In some embodiments, the methods can include using informationobtained by analysis of the nucleic acid sample by a third party. Insome embodiments, the methods can include steps that occur at more thanone site. For example, a nucleic acid sample can be obtained from asubject at a first site, such as at a health care provider or at thesubject's home in the case of a self-testing kit. The nucleic acidsample can be analyzed at the same or a second site, for example, at alaboratory or other testing facility.

Nucleic Acids

The nucleic acids and polypeptides described herein can be used inmethods and kits of the present disclosure. In some embodiments,aptamers that specifically bind the nucleic acids and polypeptidesdescribed herein can be used in methods and kits of the presentdisclosure. As used herein, a nucleic acid can comprise adeoxyribonucleotide (DNA) or ribonucleotide (RNA), whether singular orin polymers, naturally occurring or non-naturally occurring,double-stranded or single-stranded, coding, for example a translatedgene, or non-coding, for example a regulatory region, or any fragments,derivatives, mimetics or complements thereof. In some embodiments,nucleic acids can comprise oligonucleotides, nucleotides,polynucleotides, nucleic acid sequences, genomic sequences,complementary DNA (cDNA), antisense nucleic acids, DNA regions, probes,primers, genes, regulatory regions, introns, exons, open-reading frames,binding sites, target nucleic acids and allele-specific nucleic acids.

A “probe,” as used herein, includes a nucleic acid fragment forexamining a nucleic acid in a specimen using the hybridization reactionbased on the complementarity of nucleic acid.

A “hybrid” as used herein, includes a double strand formed between anyone of the abovementioned nucleic acid, within the same type, or acrossdifferent types, including DNA-DNA, DNA-RNA, RNA-RNA or the like.

“Isolated” nucleic acids, as used herein, are separated from nucleicacids that normally flank the gene or nucleotide sequence (as in genomicsequences) and/or has been completely or partially purified from othertranscribed sequences (e.g., as in an RNA library). For example,isolated nucleic acids of the disclosure can be substantially isolatedwith respect to the complex cellular milieu in which it naturallyoccurs, or culture medium when produced by recombinant techniques, orchemical precursors or other chemicals when chemically synthesized. Insome instances, the isolated material can form part of a composition,for example, a crude extract containing other substances, buffer systemor reagent mix. In some embodiments, the material can be purified toessential homogeneity using methods known in the art, for example, bypolyacrylamide gel electrophoresis (PAGE) or column chromatography(e.g., HPLC). With regard to genomic DNA (gDNA), the term “isolated”also can refer to nucleic acids that are separated from the chromosomewith which the genomic DNA is naturally associated. For example, theisolated nucleic acid molecule can contain less than about 250 kb, 200kb, 150 kb, 100 kb, 75 kb, 50 kb, 25 kb, 10 kb, 5 kb, 4 kb, 3 kb, 2 kb,1 kb, 0.5 kb or 0.1 kb of the nucleotides that flank the nucleic acidmolecule in the gDNA of the cell from which the nucleic acid molecule isderived.

Nucleic acids can be fused to other coding or regulatory sequences canbe considered isolated. For example, recombinant DNA contained in avector is included in the definition of “isolated” as used herein. Insome embodiments, isolated nucleic acids can include recombinant DNAmolecules in heterologous host cells or heterologous organisms, as wellas partially or substantially purified DNA molecules in solution.Isolated nucleic acids also encompass in vivo and in vitro RNAtranscripts of the DNA molecules of the present disclosure. An isolatednucleic acid molecule or nucleotide sequence can be synthesizedchemically or by recombinant means. Such isolated nucleotide sequencescan be useful, for example, in the manufacture of the encodedpolypeptide, as probes for isolating homologous sequences (e.g., fromother mammalian species), for gene mapping (e.g., by in situhybridization with chromosomes), or for detecting expression of thegene, in tissue (e.g., human tissue), such as by Northern blot analysisor other hybridization techniques disclosed herein. The disclosure alsopertains to nucleic acid sequences that hybridize under high stringencyhybridization conditions, such as for selective hybridization, to anucleotide sequence described herein Such nucleic acid sequences can bedetected and/or isolated by allele- or sequence-specific hybridization(e.g., under high stringency conditions). Stringency conditions andmethods for nucleic acid hybridizations are well known to the skilledperson (see, e.g., Current Protocols in Molecular Biology, Ausubel. F.et al., John Wiley & Sons, (1998), and Kraus, M. and Aaronson. S.,Methods Enzymol., 200:546-556 (1991), the entire teachings of which areincorporated by reference herein.

Calculations of “identity” or “percent identity” between two or morenucleotide or amino acid sequences can be determined by aligning thesequences for optimal comparison purposes (e.g., gaps can be introducedin the sequence of a first sequence). The nucleotides at correspondingpositions are then compared, and the percent identity between the twosequences is a function of the number of identical positions shared bythe sequences (i.e. % identity=# of identical positions/total # ofpositions×100). For example, a position in the first sequence isoccupied by the same nucleotide as the corresponding position in thesecond sequence, then the molecules are identical at that position. Thepercent identity between the two sequences is a function of the numberof identical positions shared by the sequences, taking into account thenumber of gaps, and the length of each gap, which need to be introducedfor optimal alignment of the two sequences.

In some embodiments, the length of a sequence aligned for comparisonpurposes is at least 30%, at least 40%, at least 50%, at least 60%, atleast 70%, at least 80%, at least 90%, or at least 95%, of the length ofthe reference sequence. The actual comparison of the two sequences canbe accomplished by well-known methods, for example, using a mathematicalalgorithm. A non-limiting example of such a mathematical algorithm isdescribed in Karlin, S. and Altschul, S., Proc. Natl. Acad. Sci. USA,90-5873-5877 (1993). Such an algorithm is incorporated into the NBLASTand XBLAST programs (version 2.0), as described in Altschul, S. et al.,Nucleic Acids Res., 25:3389-3402 (1997). When utilizing BLAST and GappedBLAST programs, any relevant parameters of the respective programs(e.g., NBLAST) can be used. For example, parameters for sequencecomparison can be set at score=100, word length=12, or can be varied(e.g., W=5 or W=20). Other examples include the algorithm of Myers andMiller, CABIOS (1989). ADVANCE, ADAM, BLAT, and FASTA. In someembodiments, the percent identity between two amino acid sequences canbe accomplished using, for example, the GAP program in the GCG softwarepackage (Accelrys, Cambridge, UK).

“Probes” or “primers” can be oligonucleotides that hybridize in abase-specific manner to a complementary strand of a nucleic acidmolecule. Probes can include primers, which can be a single-strandedoligonucleotide probe that can act as a point of initiation oftemplate-directed DNA synthesis using methods including but not limitedto, polymerase chain reaction (PCR) and ligase chain reaction (LCR) foramplification of a target sequence. Oligonucleotides, as describedherein, can include segments or fragments of nucleic acid sequences, ortheir complements. In some embodiments, DNA segments can be between 5and 10,000 contiguous bases, and can range from 5, 10, 12, 15, 20, or 25nucleotides to 10, 15, 20, 25, 30, 40, 50, 100, 200, 500, 1000 or 10,000nucleotides. In addition to DNA and RNA, probes and primers can includepolypeptide nucleic acids (PNA), as described in Nielsen, P. et al.,Science 254: 1497-1500 (1991). A probe or primer can comprise a regionof nucleotide sequence that hybridizes to at least about 15, typicallyabout 20-25, and in certain embodiments about 40, 50, 60 or 75,consecutive nucleotides of a nucleic acid molecule.

The present disclosure also provides isolated nucleic acids, forexample, probes or primers, that contain a fragment or portion that canselectively hybridize to a nucleic acid that comprises, or consists of,a nucleotide sequence, wherein the nucleotide sequence can comprise atleast one polymorphism or polymorphic allele contained in the geneticvariations described herein or the wild-type nucleotide that is locatedat the same position, or the complements thereof. In some embodiments,the probe or primer can be at least 70% identical, at least 80%identical, at least 85% identical, at least 90% identical, or at least95% identical, to the contiguous nucleotide sequence or to thecomplement of the contiguous nucleotide sequence.

In some embodiments, a nucleic acid probe can be an oligonucleotidecapable of hybridizing with a complementary region of a gene associatedwith a condition (e.g., LHON) containing a genetic variation describedherein. The nucleic acid fragments of the disclosure can be used asprobes or primers in assays such as those described herein.

The nucleic acids of the disclosure, such as those described above, canbe identified and isolated using standard molecular biology techniqueswell known to the skilled person. In some embodiments, DNA can beamplified and/or can be labeled (e.g., radiolabeled, fluorescentlylabeled) and used as a probe for screening, for example, a cDNA libraryderived from an organism. cDNA can be derived from mRNA and can becontained in a suitable vector. For example, corresponding clones can beisolated, DNA obtained fallowing in vivo excision, and the cloned insertcan be sequenced in either or both orientations by art-recognizedmethods to identify the correct reading frame encoding a polypeptide ofthe appropriate molecular weight. Using these or similar methods, thepolypeptide and the DNA encoding the polypeptide can be isolated,sequenced and further characterized.

In some embodiments, nucleic acid can comprise one or morepolymorphisms, variations, or mutations, for example, single nucleotidepolymorphisms (SNPs), single nucleotide variations (SNVs), copy numbervariations (CNVs), for example, insertions, deletions, inversions, andtranslocations. In some embodiments, nucleic acids can comprise analogs,for example, phosphorothioates, phosphoramidates, methyl phosphonate,chiralmethyl phosphonates, 2-O-methyl ribonucleotides, or modifiednucleic acids, for example, modified backbone residues or linkages, ornucleic acids combined with carbohydrates, lipids, polypeptide or othermaterials, or peptide nucleic acids (PNAs), for example, chromatin,ribosomes, and transcriptosomes. In some embodiments nucleic acids cancomprise nucleic acids in various structures, for example, A DNA, B DNA,Z-form DNA, siRNA, tRNA, and ribozymes. In some embodiments, the nucleicacid may be naturally or non-naturally polymorphic, for example, havingone or more sequence differences, for example, additions, deletionsand/or substitutions, as compared to a reference sequence. In someembodiments, a reference sequence can be based on publicly availableinformation, for example, the U.C. Santa Cruz Human Genome BrowserGateway (genome.ucsc.edu/cgi-bin/hgGateway) or the NCBI website(www.ncbi.nlm.nih.gov). In some embodiments, a reference sequence can bedetermined by a practitioner of the present disclosure using methodswell known in the art, for example, by sequencing a reference nucleicacid.

In some embodiments, a probe can hybridize to an allele, SNP, SNV, orCNV as described herein. In some embodiments, the probe can bind toanother marker sequence associated with LHON as described herein.

One of skill in the art would know how to design a probe so thatsequence specific hybridization can occur only if a particular allele ispresent in a genomic sequence from a test nucleic acid sample. Thedisclosure can also be reduced to practice using any convenientgenotyping method, including commercially available technologies andmethods for genotyping particular genetic variations

Control probes can also be used, for example, a probe that binds a lessvariable sequence, for example, a repetitive DNA associated with acentromere of a chromosome, can be used as a control. In someembodiments, probes can be obtained from commercial sources. In someembodiments, probes can be synthesized, for example, chemically or invitro, or made from chromosomal or genomic DNA through standardtechniques. In some embodiments sources of DNA that can be used includegenomic DNA, cloned DNA sequences, somatic cell hybrids that containone, or a part of one, human chromosome along with the normal chromosomecomplement of the host, and chromosomes purified by flow cytometry ormicrodissection. The region of interest can be isolated through cloning,or by site-specific amplification using PCR.

One or more nucleic acids for example, a probe or primer, can also belabeled, for example, by direct labeling, to comprise a detectablelabel. A detectable label can comprise any label capable of detection bya physical, chemical, or a biological process for example, a radioactivelabel, such as 32P or 3H, a fluorescent label, such as FITC, achromophore label, an afinity-ligand label, an enzyme label, such asalkaline phosphatase, horseradish peroxidase, or 12 galactosidase, anenzyme cofactor label, a hapten conjugate label, such as digoxigenin ordinitrophenyl, a Raman signal generating label, a magnetic label, a spinlabel, an epitope label, such as the FLAG or HA epitope, a luminescentlabel, a heavy atom label, a nanoparticle label, an electrochemicallabel, a light scattering label, a spherical shell label, semiconductornanocrystal label, such as quantum dots (described in U.S. Pat. No.6,207,392), and probes labeled with any other signal generating labelknown to those of skill in the art, wherein a label can allow the probeto be visualized with or without a secondary detection molecule. Anucleotide can be directly incorporated into a probe with standardtechniques, for example, nick translation, random priming, and PCRlabeling. A “signal,” as used herein, include a signal suitablydetectable and measurable by appropriate means, including fluorescence,radioactivity, chemiluminescence, and the like.

Non-limiting examples of label moieties useful for detection include,without limitation, suitable enzymes such as horseradish peroxidase,alkaline phosphatase, beta-galactosidase, or acetylcholinesterase;members of a binding pair that are capable of forming complexes such asstreptavidin/biotin, avidin/biotin or an antigen/antibody complexincluding, for example, rabbit IgG and anti-rabbit IgG; fluorophoressuch as umbelliferone, fluorescein, fluorescein isothiocyanate,rhodamine, tetramethyl rhodamine, cosin, green fluorescent protein,erythrosin, coumarin, methyl coumarin, pyrene, malachite green,stilbene, lucifer yellow, Cascade Blue, Texas Red,dichlorotriazinylamine fluorescein, dansyl chloride, phycoerythrin,fluorescent lanthanide complexes such as those including Europium andTerbium, cyanine dye family members, such as Cy3 and Cy5, molecularbeacons and fluorescent derivatives thereof, as well as others known inthe art as described, for example, in Principles of FluorescenceSpectroscopy, Joseph R. Lakowicz (Editor), Plenum Pub Corp, 2nd edition(July 1999) and the 6th Edition of the Molecular Probes Handbook byRichard P. Hoagland; a luminescent material such as luminol; lightscattering or plasmon resonant materials such as gold or silverparticles or quantum dots; or radioactive material include 14C, 123I,124I, 125I, Tc99m, 32P, 33P, 35S or 3H.

I Other labels can also be used in the methods of the presentdisclosure, for example, backbone labels. Backbone labels comprisenucleic acid stains that bind nucleic acids in a sequence independentmanner. Non-limiting examples include intercalating dyes such asphenanthridines and acridines (e.g., ethidium bromide, propidium iodide,hexidium iodide, dihydroethidium, ethidium homodimer-1 and -2, ethidiummonoazide, and ACMA); some minor grove binders such as indoles andimidazoles (e.g., Hoechst 33258, Hoechst 33342, Hoechst 34580 and DAPI);and miscellaneous nucleic acid stains such as acridine orange (alsocapable of intercalating), 7-AAD, actinomycin D, LDS751, andhydroxystilbamidine. All of the aforementioned nucleic acid stains arecommercially available from suppliers such as Molecular Probes, Inc.Still other examples of nucleic acid stains include the following dyesfrom Molecular Probes: cyanine dyes such as SYTOX Blue, SYTOX Green,SYTOX Orange. POPO-1, POPO-3, YOYO-1, YOYO-3, TOTO-1, TOTO-3, JOJO-1,LOLO-1, BOBO-1, BOBO-3, PO-PRO-1, PO-PRO-3, BO-PRO-1, BO-PRO-3,TO-PRO-1, TO-PRO-3, TO-PRO-5, JO-PRO-1, LO-PRO-1, YO-PRO-1, YO-PRO-3,PicoGreen, OliGreen, RiboGreen, SYBR Gold, SYBR Green I, SYBR Green II,SYBR DX, SYTO-40, -41, -42, -43, -44, -45 (blue), SYTO-13, -16, -24,-21, -23, -12, -11, -20, -22, -15, -14, -25 (green), SYTO-81, -80, -82,-83, -84, -85 (orange), SYTO-64, -17, -59, -61, -62, -60, -63 (red).

In some embodiments, fluorophores of different colors can be chosen, forexample, 7-amino-4-methylcoumarin-3-acetic acid (AMCA),5-(and-6)-carboxy-X-rhodamine, lissamine rhodamine B,5-(and-6)-carboxyfluorescein, fluoreseein-5-isothiocyanate (FITC),7-diethylaminocoumarin-3-carboxvlic acid,tetramethvlrhodamine-5-(and-6)-isothiocvanate,5-(and-6)-carboxytetramethylrhodamine, 7-hydroxycoumarin-3-carboxylicacid, 6-[fluorescein 5-(and-6)-carboxamido]hexanoic acid,N-(4,4-difluoro-5,7-dimethyl-4-bora-3a,4a diaza-3-indacenepropionicacid, cosin-5-isothiocyanate, erythrosin-5-isothiocyanate, TRITC,rhodamine, tetramethylrhodamine, R-phycoerythrin, Cy-3, Cy-5, Cy-7,Texas Red, Phar-Red, allophycocvanin (APC), and CASCADE™ blueacetylazide, such that each probe in or not in a set can be distinctlyvisualized. In some embodiments, fluorescently labeled probes can beviewed with a fluorescence microscope and an appropriate filter for eachfluorophore, or by using dual or triple band-pass filter sets to observemultiple fluorophores. In some embodiments, techniques such as flowcytometry can be used to examine the hybridization pattern of theprobes.

In other embodiments, the probes can be indirectly labeled, for example,with biotin or digoxygenin, or labeled with radioactive isotopes such as32P and/or 3H. As a non-limiting example, a probe indirectly labeledwith biotin can be detected by avidin conjugated to a detectable marker.For example, avidin can be conjugated to an enzymatic marker such asalkaline phosphatase or horseradish peroxidase. In some embodiments,enzymatic markers can be detected using colorimetric rcactions using asubstrate and/or a catalyst for the enzyme. In some embodiments,catalysts for alkaline phosphatase can be used, for example,5-bromo-4-chloro-3-indolylphosphate and nitro blue tetrazolium. In someembodiments, a catalyst can be used for horseradish peroxidase, forexample, diaminobenzoate.

Formulations, Routes of Administration, and Effective Doses

Yet another aspect of the present disclosure relates to formulations,routes of administration and effective doses for pharmaceuticalcompositions comprising an agent or combination of agents of the instantdisclosure. Such pharmaceutical compositions can be used to treat acondition (e.g., LHON) as described above.

Compounds of the disclosure can be administered as pharmaceuticalformulations including those suitable for oral (including buccal andsub-lingual), rectal, nasal, topical, transdermal patch, pulmonary,vaginal, suppository, or parenteral (including intraocular,intravitreal, intramuscular, intraarterial, intrathecal, intradermal,intraperitoneal, subcutaneous and intravenous) administration or in aform suitable for administration by aerosolization, inhalation orinsufflation. General information on drug delivery systems can be foundin Ansel et al., Pharmaceutical Dosage Forms and Drug Delivery Systems(Lippencott Williams & Wilkins, Baltimore Md. (1999).

In various embodiments, the pharmaceutical composition includes carriersand excipients (including but not limited to buffers, carbohydrates,mannitol, polypeptides, amino acids, antioxidants, bacteriostats,chelating agents, suspending agents, thickening agents and/orpreservatives), water, oils including those of petroleum, animal,vegetable or synthetic origin, such as peanut oil, soybean oil, mineraloil, sesame oil and the like, saline solutions, aqueous dextrose andglycerol solutions, flavoring agents, coloring agents, detackifiers andother acceptable additives, adjuvants, or binders, otherpharmaceutically acceptable auxiliary substances to approximatephysiological conditions, such as pH buffering agents, tonicityadjusting agents, emulsifying agents, wetting agents and the like.Examples of excipients include starch, glucose, lactose, sucrose,gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerolmonostearate, talc, sodium chloride, dried skim milk, glycerol,propylene, glycol, water, ethanol and the like. In some embodiments, thepharmaceutical preparation is substantially free of preservatives. Inother embodiments, the pharmaceutical preparation can contain at leastone preservative. General methodology on pharmaceutical dosage forms isfound in Ansel et al., Pharmaceutical Dosage Forms and Drug DeliverySystems (Lippencott, Williams, & Wilkins, Baltimore Md. (1999)). It canbe recognized that, while any suitable carrier known to those ofordinary skill in the art can be employed to administer the compositionsof this disclosure, the type of carrier can vary depending on the modeof administration.

Compounds can also be encapsulated within liposomes using well-knowntechnology. Biodegradable microspheres can also be employed as carriersfor the pharmaceutical compositions of this disclosure. Suitablebiodegradable microspheres are disclosed, for example, in U.S. Pat. Nos.4,897,268, 5,075,109, 5,928,647, 5,811,128, 5,820,883, 5,853,763,5,814,344 and 5,942,252.

The compound can be administered in liposomes or microspheres (ormicroparticles). Methods for preparing liposomes and microspheres foradministration to a subject are well known to those of skill in the art.U.S. Pat. No. 4,789,734, the contents of which are hereby incorporatedby reference, describes methods for encapsulating biological materialsin liposomes. Essentially, the material is dissolved in an aqueoussolution, the appropriate phospholipids and lipids added, and along withsurfactants if required, and the material dialyzed or sonicated, asnecessary. A review of known methods is provided by G. Gregoriadis,Chapter 14, “Liposomes.” Drug Carriers in Biology and Medicine, pp.2.sup.87-341 (Academic Press, 1979).

Microspheres formed of polymers or polypeptides are well known to thoseskilled in the art, and can be tailored for passage through thegastrointestinal tract directly into the blood stream. Alternatively,the compound can be incorporated and the microspheres, or composite ofmicrospheres, implanted for slow release over a period of time rangingfrom days to months. See, for example, U.S. Pat. Nos. 4,906,474,4,925,673 and 3,625,214, and Jein, TIPS 19:155-157 (1998), the contentsof which are hereby incorporated by reference.

The concentration of drug can be adjusted, the pH of the solutionbuffered and the isotonicity adjusted to be compatible with intraocularor intravitreal injection.

The compounds of the disclosure can be formulated as a sterile solutionor suspension, in suitable vehicles. The pharmaceutical compositions canbe sterilized by conventional, well-known sterilization techniques, orcan be sterile filtered. The resulting aqueous solutions can be packagedfor use as is, or lyophilized, the lyophilized preparation beingcombined with a sterile solution prior to administration. Suitableformulations and additional carriers are described in Remington “TheScience and Practice of Pharmacy” (20th Ed., Lippincott Williams &Wilkins, Baltimore Md.), the teachings of which are incorporated byreference in their entirety herein.

The agents or their pharmaceutically acceptable salts can be providedalone or in combination with one or more other agents or with one ormore other forms. For example, a formulation can comprise one or moreagents in particular proportions, depending on the relative potencies ofeach agent and the intended indication. For example, in compositions fortargeting two different host targets, and where potencies are similar,about a 1:1 ratio of agents can be used. The two forms can be formulatedtogether, in the same dosage unit e.g., in one cream, suppository,tablet, capsule, aerosol spray, or packet of powder to be dissolved in abeverage; or each form can be formulated in a separate unit, e.g., twocreams, two suppositories, two tablets, two capsules, a tablet and aliquid for dissolving the tablet, two aerosol sprays, or a packet ofpowder and a liquid for dissolving the powder, etc.

The term “pharmaceutically acceptable salt” means those salts whichretain the biological effectiveness and properties of the agents used inthe present disclosure, and which are not biologically or otherwiseundesirable.

Typical salts are those of the inorganic ions, such as, for example,sodium, potassium, calcium, magnesium ions, and the like. Such saltsinclude salts with inorganic or organic acids, such as hydrochloricacid, hydrobromic acid, phosphoric acid, nitric acid, sulfuric acid,methanesulfonic acid, p toluenesulfonic acid, acetic acid, fumaric acid,succinic acid, lactic acid, mandelic acid, malic acid, citric acid,tartaric acid or maleic acid. In addition, if the agent(s) contain acarboxyl group or other acidic group, it can be converted into apharmaceutically acceptable addition salt with inorganic or organicbases. Examples of suitable bases include sodium hydroxide, potassiumhydroxide, ammonia, cyclohexylamine, dicyclohexyl-amine, ethanolamine,diethanolamine, triethanolamine, and the like.

A pharmaceutically acceptable ester or amide refers to those whichretain biological effectiveness and properties of the agents used in thepresent disclosure, and which are not biologically or otherwiseundesirable. Typical esters include ethyl, methyl, isobutyl, ethyleneglycol, and the like. Typical amides include unsubstituted amides, alkylamides, dialkyl amides, and the like.

In some embodiments, an agent can be administered in combination withone or more other compounds, forms, and/or agents. e.g., as describedabove. Pharmaceutical compositions with one or more other active agentscan be formulated to comprise certain molar ratios. For example, molarratios of about 99:1 to about 1:99 of a first active agent to the otheractive agent can be used. In some subset of the embodiments, the rangeof molar ratios of a first active agent: other active agents areselected from about 80:20 to about 20:80; about 75:25 to about 25:75,about 70:30 to about 30:70, about 66:33 to about 33:66, about 60:40 toabout 40:60; about 50:50; and about 90:10 to about 10:90. The molarratio of a first active: other active agents can be about 1:9, and insome embodiments can be about 1:1. The two agents, forms and/orcompounds can be formulated together, in the same dosage unit e.g., inone cream, suppository, tablet, capsule, or packet of powder to bedissolved in a beverage; or each agent, form, and/or compound can beformulated in separate units, e.g., two creams, suppositories, tablets,two capsules, a tablet and a liquid for dissolving the tablet, anaerosol spray a packet of powder and a liquid for dissolving the powder,etc.

If necessary or desirable, the agents and/or combinations of agents canbe administered with still other agents. The choice of agents that canbe co-administered with the agents and/or combinations of agents of theinstant disclosure can depend, at least in part, on the condition beingtreated.

The agent(s) (or pharmaceutically acceptable salts, esters or amidesthereof) can be administered per se or in the form of a pharmaceuticalcomposition wherein the active agent(s) is in an admixture or mixturewith one or more pharmaceutically acceptable carriers. A pharmaceuticalcomposition, as used herein, can be any composition prepared foradministration to a subject. Pharmaceutical compositions for use inaccordance with the present disclosure can be formulated in conventionalmanner using one or more physiologically acceptable carriers, comprisingexcipients, diluents, and/or auxiliaries, e.g., which facilitateprocessing of the active agents into preparations that can beadministered. Proper formulation can depend at least in part upon theroute of administration chosen. The agent(s) useful in the presentdisclosure, or pharmaceutically acceptable salts, esters, or amidesthereof, can be delivered to a subject using a number of routes or modesof administration, including oral, buccal, topical, rectal, transdermal,transmucosal, subcutaneous, intravenous, intraocular, intravitreal, andintramuscular applications, as well as by inhalation.

In some embodiments, oils or non-aqueous solvents can be used to bringthe agents into solution, due to, for example, the presence of largelipophilic moieties. Alternatively, emulsions, suspensions, or otherpreparations, for example, liposomal preparations, can be used. Withrespect to liposomal preparations, any known methods for preparingliposomes for treatment of a condition can be used. See, for example,Bangham et al., J. Mol. Biol. 23: 238-252 (1965) and Szoka et al., Proc.Natl Acad. Sci. USA 75: 4194-4198 (1978), incorporated herein byreference. Ligands can also be attached to the liposomes to direct thesecompositions to particular sites of action. Agents of this disclosurecan also be integrated into foodstuffs, e.g., cream cheese, butter,salad dressing, or ice cream to facilitate solubilization,administration, and/or compliance in certain subject populations.

The compounds of the disclosure can be formulated for parenteraladministration (e.g., by injection, for example, intraocular orintravitreal injection) and can be presented in unit dose form inampoules, pre-filled syringes, small volume infusion or in multi-dosecontainers with an added preservative. The compositions can take suchforms as suspensions, solutions, or emulsions in oily or aqueousvehicles, for example, solutions in aqueous polyethylene glycol.

For injectable formulations, the vehicle can be chosen from those knownin art to be suitable, including aqueous solutions or oil suspensions,or emulsions, with sesame oil, corn oil, cottonseed oil, or peanut oil,as well as elixirs, mannitol, dextrose, or a sterile aqueous solution,and similar pharmaceutical vehicles. The formulation can also comprisepolymer compositions which are biocompatible, biodegradable, such aspoly(lactic-co-glycolic)acid. These materials can be made into micro ornanospheres, loaded with drug and further coated or derivatized toprovide superior sustained release performance. Vehicles suitable forperiocular or intraocular injection include, for example, suspensions oftherapeutic agent in injection grade water, liposomes and vehiclessuitable for lipophilic substances. Other vehicles for periocular orintraocular injection are well known in the art.

In some embodiments, the composition is formulated in accordance withroutine procedures as a pharmaceutical composition adapted forintravenous administration to human beings. Typically, compositions forintravenous administration are solutions in sterile isotonic aqueousbuffer. Where necessary, the composition can also include a solubilizingagent and a local anesthetic such as lidocaine to ease pain at the siteof the injection. Generally, the ingredients are supplied eitherseparately or mixed together in unit dosage form, for example, as a drylyophilized powder or water free concentrate in a hermetically sealedcontainer such as an ampoule or sachette indicating the quantity ofactive agent. Where the composition is to be administered by infusion,it can be dispensed with an infusion bottle containing sterilepharmaceutical grade water or saline. Where the composition isadministered by injection, an ampoule of sterile water for injection orsaline can be provided so that the ingredients can be mixed prior toadministration.

When administration is by injection, the active compound can beformulated in aqueous solutions, specifically in physiologicallycompatible buffers such as Hanks solution, Ringer's solution, orphysiological saline buffer. The solution can contain formulatory agentssuch as suspending, stabilizing and/or dispersing agents. Alternatively,the active compound can be in powder form for constitution with asuitable vehicle, e.g., sterile pyrogen-free water, before use. In someembodiments, the pharmaceutical composition does not comprise anadjuvant or any other substance added to enhance the immune responsestimulated by the peptide. In some embodiments, the pharmaceuticalcomposition comprises a substance that inhibits an immune response tothe peptide. Methods of formulation are known in the art, for example,as disclosed in Remington's Pharmaceutical Sciences, latest edition,Mack Publishing Co., Easton P.

In some embodiments, eye disorders can be effectively treated withophthalmic solutions, suspensions, ointments or inserts comprising anagent or combination of agents of the present disclosure. Eye drops canbe prepared by dissolving the active ingredient in a sterile aqueoussolution such as physiological saline, buffering solution, etc., or bycombining powder compositions to be dissolved before use. Other vehiclescan be chosen, as is known in the art, including but not limited to:balance salt solution, saline solution, water soluble polyethers such aspolyethyene glycol, polyvinyls, such as polyvinyl alcohol and povidone,cellulose derivatives such as methylcellulose and hydroxypropylmethylcellulose, petroleum derivatives such as mineral oil and whitepetrolatum, animal fats such as lanolin, polymers of acrylic acid suchas carboxypolymethylene gel, vegetable fats such as peanut oil andpolysaccharides such as dextrans, and glycosaminoglycans such as sodiumhyaluronate. If desired, additives ordinarily used in the eye drops canbe added. Such additives include isotonizing agents (e.g., sodiumchloride, etc.), buffer agent (e.g., boric acid, sodium monohydrogenphosphate, sodium dihydrogen phosphate, etc.), preservatives (e.g.,benzalkonium chloride, benzethonium chloride, chlorobutanol, etc.),thickeners (e.g., saccharide such as lactose, mannitol, maltose, etc.;e.g., hyaluronic acid or its salt such as sodium hyaluronate, potassiumhyaluronate, etc.; e.g., mucopolysaccharide such as chondroitin sulfate,etc.; e.g., sodium polyacrylate, carboxyvinyl polymer, crosslinkedpolyacrylate, polyvinyl alcohol, polyvinyl pyrrolidone, methylcellulose, hydroxy propyl methylcellulose, hydroxyethyl cellulose,carboxymethyl cellulose, hydroxy propyl cellulose or other agents knownto those skilled in the art).

The solubility of the components of the present compositions can beenhanced by a surfactant or other appropriate co-solvent in thecomposition. Such cosolvents include polysorbate 20, 60, and 80,Pluronic F68, F-84 and P-103, cyclodextrin, or other agents known tothose skilled in the art. Such co-solvents can be employed at a level offrom about 0.01% to 2% by weight.

The compositions of the disclosure can be packaged in multidose form.Preservatives can be preferred to prevent microbial contamination duringuse. Suitable preservatives include: benzalkonium chloride, thimerosal,chlorobutanol, methyl paraben, propyl paraben, phenylethyl alcohol,edetate disodium, sorbic acid, Onamer M. or other agents known to thoseskilled in the art. In the prior art ophthalmic products, suchpreservatives can be employed at a level of from 0.004% to 0.02%. In thecompositions of the present application the preservative, preferablybenzalkonium chloride, can be employed at a level of from 0.001% to lessthan 0.01%, e.g., from 0.001% to 0.008%, preferably about 0.005% byweight. It has been found that a concentration of benzalkonium chlorideof 0.005% can be sufficient to preserve the compositions of the presentdisclosure from microbial attack.

In some embodiments, the agents of the present disclosure are deliveredin soluble rather than suspension form, which allows for more rapid andquantitative absorption to the sites of action. In general, formulationssuch as jellies, creams, lotions, suppositories and ointments canprovide an area with more extended exposure to the agents of the presentdisclosure, while formulations in solution, e.g., sprays, provide moreimmediate, short-term exposure.

It is envisioned additionally, that the compounds of the disclosure canbe attached releasably to biocompatible polymers for use in sustainedrelease formulations on, in or attached to inserts for topical,intraocular, periocular, or systemic administration. The controlledrelease from a biocompatible polymer can be utilized with a watersoluble polymer to form an instillable formulation, as well. Thecontrolled release from a biocompatible polymer, such as for example,PLGA microspheres or nanospheres, can be utilized in a formulationsuitable for intra ocular implantation or injection for sustainedrelease administration, as well any suitable biodegradable andbiocompatible polymer can be used.

EXAMPLES

The following exemplary embodiments further describe the presentinvention. It should be understood that these examples are only intendedto illustrate the invention, but not to limit the scope of the presentinvention. Unless otherwise indicated, the methods and conditionsdisclosed in e.g., sambrook et al, molecular cloning: a laboratorymanual (New York: cold spring harbor laboratory press, 1989) or theconditions recommended by the manufacturer can be used in the examplesbelow.

Example 1—ND4 Plasmid and Virus Preparation

1.1 Plasmid Preparation

The nucleotide sequence for human ND4 (SEQ ID NO: 6) was obtained basedon US National Center for Biotechnology Information reference sequenceyp_003024035.1. The sequences for the non-optimized mitochondrialtargeting sequence COX10 is SEQ ID NO: 1. The optimized sequences forthe mitochondrial targeting sequence COX10 (opt_COX10, SEQ ID NO: 2) andthe coding sequence of human ND4 (opt_ND4, SEQ ID NO: 7) were designedto improve the transcription efficiency and the translation efficiency.The optimized COX10-ND4 sequence, which is about 75.89% homology to thenon-optimized COX10-ND4, was followed by a three prime untranslatedregion (i.e., 3′UTR, SEQ ID NO: 13) to a recombinant nucleic acid,opt_COX10-opt_ND4-3′UTR (as shown in SEQ ID NO: 31).

The synthesized recombinant nucleic acid, opt_COX10-opt_ND4-3′UTR, wasincorporated into an adeno-associated virus (AAV) vector by PCRamplification (FIG. 1). The opt_COX10-opt_ND4-3′UTR was cut by theEcoRI/SalI restriction enzymes to form cohesive ends, and then embeddedinto an AAV vector with EcoRI/SalI restriction sites, such as the pSNaVvector, to generate the pSNaV/rAAV2/2-ND4 plasmid (i.e., thepAAV2-optimized ND4 plasmid). The pAAV2-opt_ND4 plasmid was compared tothe non-optimized pAAV2-ND4 plasmid.

The recon screening and identifying steps were similar to theCN102634527B: the plasmid was cultured at 37° C. in a LB plate. Bluecolonies and white colonies were appeared, where white colonies wererecombinant clones. The white colonies were picked, added to 100 mg/Lampicillin-containing LB culture medium, cultured at 37° C., 200 rpm for8 hours and then the plasmid were extracted from the cultured bacterialmedium based on the Biomiga plasmid extraction protocol. Theidentification of the plasmid was confirmed using the EcoRI/SalIrestriction enzymes.

1.2 Cell Transfection

One day before transfection, HEK293 cells were inoculated to 225 cm²cell culture bottle: at the inoculation density of 3.0×10⁷ cells/ml, theculture medium was the Dulbecco's Modified Eagle Medium (DMEM) with 10%bovine serum, at 37° C. in a 5% CO2 incubator overnight. The culturemedium were replaced with fresh DMEM with 10% bovine serum on the day oftransfection.

After the cells grow to 80-90%, discard the culture medium and transfectthe cells with the pAAV2-ND4 and pAAV2-opt_ND4 plasmid, using thePlasmidTrans (VGTC) transfection kit. The detailed transfection protocolwas described in CN102634527B example 1. The cells were collected 48 hafter the transfection.

1.3 Collection, Concentration and Purification of the RecombinantAdeno-Associated Virus

Virus collection: 1) dry ice ethanol bath (or liquid nitrogen) and a 37°C. water bath were prepared; 2) the transfected cells along with mediawere collected in a 15 ml centrifuge tube, 3) the cells were centrifugedfor 3 minutes at 1000 rpm/min; the cells and supernatant were separated,the supernatant were stored separately; and the cells were re-suspendedin 1 ml of PBS: 4) the cell suspension were transferred between the dryice-ethanol bath and 37° C. water bath repeatedly, freeze thawing forfour times for 10 minutes each, slightly shaking after each thawing.

Virus concentration: 1) cell debris were removed with 10,000 gcentrifugation; the centrifugal supernatant was transferred to a newcentrifuge tube; 2) impurities were removed by filtering with a 0.45 μmfilter; 3) each ½ volume of 1M NaCl and 10% PEG 8000 solution were addedin the sample, uniformly mixed, and stored at 4° C. overnight; 4)supernatant was discarded after 12,000 rpm centrifugation for 2 h; afterthe virus precipitate was completely dissolving in an appropriate amountof PBS solution, sterilizing the sample with a 0.22 μm filter; 5) addingbenzonase nuclease was added to remove residual plasmid DNA (finalconcentration at 50 U/ml). The tube was inverted several times to mixthoroughly and then incubated at 37° C. for 30 minutes; 6) the samplewas filtered with a 0.45 μm filtration head; the filtrate is theconcentrated rAAV2 virus.

Virus purification: 1) CsCl was added to the concentrated virus solutionuntil a density of 1.41 g/ml (refraction index at 1.372): 2) the samplewas added to in the ultracentrifuge tube and filled the tube withpre-prepared 1.41 g/ml CsCl solution: 3) centrifuged at 175,000 g for 24hours to form a density gradient. Sequential collection of differentdensities of the sample was performed. The enriched rAAV2 particles werecollected; 4) repeating the process one more time. The virus was loadedto a 100 kDa dialysis bag and dialyzed/desalted at 4° C. overnight. Theconcentrated and purified recombinant adeno-associated virus wererAAV2-ND4 and rAAV2-optimized ND4.

Similarly, other mitochondrial targeting sequences (MTS), such as OPA1(SEQ ID NO: 5) can be used to replace COX10 in the above example andcreate AAV with recombinant plasmids.

Example 2—Intravitreal Injection of rAAV2 in Rabbit Eyes

Twelve rabbits were divided into 2 group: rAAV2-ND4 and rAAV2-optimizedND4. Virus solution (1-10¹ vg/0.05 mL) was punctured into the vitreouscavity from 3 mm outside the corneal limbus at the pars plana. After theintravitreal injection, the eyes were examined using slit lamp exam andfundus photography inspection. Injection for 30 days. RT-PCR detectionand immunoblotting were carried out in each group respectively.

Example 3—Real-Time PCR for the Expression of ND4

The RNAs from the transfected rAAV2-ND4 and rAAV2-optimized ND4 rabbitoptic nerve cells were extracted using the TRIZOL total RNA extractionkit. cDNA templates were synthesized by reverse transcription of theextracted RNA.

The NCBI conserved structural domain analysis software were used toanalyze the conservative structure of ND4, ensuring that the designedprimers amplified fragments were located at non-conserved region; thenprimers were designed according to the fluorescent quantitative PCRprimer design principle:

β-actin-S: (SEQ ID NO: 85) CGAGATCGTGCGGGACAT; β-actin-A:(SEQ ID NO: 86) CAGGAAGGAGGGCTGGAAC; ND4-S: (SEQ ID NO: 87)CTGCCTACGACAAACAGAC; ND4-A: (SEQ ID NO: 88) AGTGCGTTCGTAGTTTGAG;

The fluorescent quantitative PCR reaction and protocol: fluorescencequantitative PCR were measured in a real-time PCR detection system. In a0.2 ml PCR reaction tube, SYBR green mix 12.5 W, ddH₂O 8 μl, 1 sW ofeach primer, and the cDNA sample 2.5 μl, were added to an overall volumeof 25 μl. Each sample was used for amplification of the target gene andamplifying the reference gene β-actin, and each amplification wererepeated three times. The common reagents were added together and thendivided separately to minimize handling variation. The fluorescentquantitative PCR were carried out: pre-denaturation at 95° C. for 1 s,denaturation at 94° C. for 15 s, annealing at 55° C. for 15 sec,extension at 72° C. for 45 s. A total of 40 cycles of amplificationreaction were performed and fluorescence signal acquisition was done atthe extension phase of each cycle. After the reaction, a 94° C. to 55°C. melting curve analysis was done. By adopting a relative quantitativemethod research of gene expression level difference to beta-actin wasused as an internal reference gene.

As shown in FIG. 2, the relative expression level (mRNA level) of therAAV2-ND4 and rAAV2-optimized ND4 were 0.42±0.23 and 0.57±0.62,respectively (p<0.05, FIG. 2). The results unexpectedly show that theoptimized ND4 (opt_ND4, SEQ ID NO: 7) coding nucleic acid sequence andthe corresponding recombinant nucleic acid (opt_COX10-opt_ND4-3′UTR, SEQID NO: 31) surprisingly increased the transcription efficiency,increasing the expression of the rAAV2-optimized ND4 by about 36%. Theresults showed that the transcription efficiency of the rAAV2-optimizedND4 is significantly higher.

Example 4—Immunoblotting Detection of ND4 Expression

The ND4 protein was purified from the rabbit nerve cells transfected byrAAV2-optimized ND4 and rAAV2-ND4, respectively. After a 10%polyacrylamide gel electrophoresis, and transferred to a polyvinylidenedifluoride membrane (Bio-Rad, HER-hercules, CA, USA) for immunedetection. β-actin was used as an internal reference gene. The filmstrip was observed on an automatic image analysis instrument (Li-Cor;Lincoln, Nebr., USA) and analyzed using the integrated optical densityof the protein band with integral normalization method, so as to obtainthe same sample corresponding optical density value. The statisticalanalysis software SPSS 19.0 was used for the data analysis.

The results was shown in FIG. 3. The average relative protein expressionlevel of ND4 for rAAV2-optimized ND4 (left black column) and rAAV2-ND4was 0.32±0.11 and 0.68 t 0.20, respectively (p<0.01, FIG. 3). Theresults unexpectedly show that the optimized ND4 coding nucleic acidsequence (opt_ND4, SEQ ID NO: 7) and the corresponding recombinantnucleic acid (opt_COX10-opt_ND4-3′UTR, SEQ ID NO: 31) surprisinglyincreased the translation efficiency, increasing the expression of therAAV2-optimized ND4 by about 112%. The results showed that thetranslation efficiency of the rAAV2-optimized ND4 is also significantlyhigher.

Example 5—Rabbits Intraocular Pressure and Eye-Ground Photography

Slit lamp examination and intraocular pressure measurement was performedon both groups of rabbits at 1, 3, 7, and 30 days after the surgery. Noobviously abnormality, conjunctival congestion, secretions, orendophthalmitis were observed and the intraocular pressure were notelevated in all the rabbits.

The fundus photographic results were shown in FIG. 4. No obvious damageor complication to the optic nerve and retinal vascular of the rabbits,indicating the standard intravitreal injection is safe withoutnoticeable inflammation reaction or other complications.

Example 6—Human Clinical Trial

Two groups of patients were tested: 1) between 2011 and 2012, 9 patientsreceived intravitreal injection of 1-10¹⁰ vg/0.05 mL rAAV2-ND4 in asingle eye, as a control group; and 2) between 2017 and January 2018, 20patients received intravitreal injection of 1×10¹⁰ vg/0.05 mLrAAV2-optimized ND4 in a single eye, as an experimental group. Theresults of the clinical trial were analyzed using the statisticalanalysis SPSS 19.0.

The comparison of the two groups is shown in Table 2. The fastesteyesight improving time was 1 month in the experimental group, which wassignificantly faster than the control group at 3 months (p<0.01): theoptimal recovery of vision for the experimental group was 1.0, which wasobviously higher than the control group at 0.8 (p<0.01); the averagerecovery of vision in the experimental group was 0.582±0.086, which wasobviously higher than the control group at 0.344±0.062 (p<0.01). Thefundus photographic results were shown in FIG. 5. No obvious damage orcomplication to the optic nerve and retinal vascular of the patients inthe experimental and control groups, indicating the safety of theintravitreal injection of rAAV2-optimized ND4 and rAAV2-ND4.

TABLE 2 The comparison of rAAV2-optimized ND4 and rAAV2-ND4 in LHON genetherapy Fastest eyesight Number of optimal average Patient improvingtime patients with recovery of recovery of group number (month) improvedvision vision vision control 9 3  6 (67%) 0.8 0.344 ± 0.062 experimental20 1 15 (75%) 1.0 0.582 ± 0.086 P value <0.01 <0.01 <0.01 <0.01

Example 7—OPA1 as the Mitochondrial Targeting Sequences

The COX10 and 3′UTR sequences in the recombinant nucleic acid(opt_COX10-opt_ND4-3′UTR, SEQ ID NO: 31) in examples 1-6 were replacedwith another mitochondrial targeted sequence, OPA1 (SEQ ID NO: 5) andanother 3′UTR sequence, 3′UTR* (SEQ ID NO: 14) respectively, to generatea new recombinant nucleic acid, OPA1-opt_ND4-3′UTR* (SEQ ID NO: 74).

Experimental methods were the same as examples 1-6, where therecombinant nucleic acid opt_COX10-opt_ND4-3′UTR (SEQ ID NO: 31) wasreplaced by OPA1-opt_ND4-3′UTR* (SEQ ID NO: 74). It was found that, theoptimized ND4 sequence has significantly improved transcription andtranslation efficiencies, expression levels, as well as higher efficacyand safety in treating LHON when compared to non-optimized ND4(COX10-ND4-3′UTR, SEQ ID NO: 15).

Example 8—Optimized ND4 Sequence Opt_ND4*

Similar experimental methods in examples 1-6 were followed using thenucleic acid, opt_COX10*-opt_ND4*-3′UTR (SEQ ID NO: 47). Follow thesimilar procedures as in example 1, virus tagged with a fluorescentprotein. EGFP, was prepared as rAAV2-ND4-EGFP and rAAV2-opt_ND4*-EGFP.

The frozen 293T cell was resuscitated and allowed to grow in a T75 flaskto about 90%. The cells were precipitated and resuspended in DMEMcomplete medium to a cell density of 5×10⁴ cells/mL. The cells wereresuspended. About 100 μl of the cell suspension (about 5000 cells) wereadded in each well of a 96 well plate. The cells were cultured and grownto 50% under 37° C. and 5% CO₂. About 0.02 μl PBS was mixed with 2×10 μlvg/0.02 μl of the virus rAAV2-ND4-EGFP and rAAV2-opt_ND4*-EGFP,respectively. After 48 hours, fluorescence microscopy and RT-PCRdetection and immunoblotting experiments were performed. As shown inFIG. 6, EGFP was successfully expressed, indicating that rAAV carryingthe EGFP gene was successfully transfected in the 293T cells andrAAV2-ND4-EGFP and rAAV2-opt_ND4*-EGFP were successfully expressed.

Real-time PCR tests similar to example 3 was following using thefollowing primers:

β-actin-S: (SEQ ID NO: 85) CGAGATCGTGCGGGACAT; β-actin-A:(SEQ ID NO: 86) CAGGAAGGAGGGCTGGAAC; ND4-S: (SEQ ID NO: 107)GCCAACAGCAACTACGAGC; ND4-A: (SEQ ID NO: 108) TGATGTTGCTCCAGCTGAAG;

The results unexpectedly show that the optimized ND4* (opt_ND4, SEQ IDNO: 8) coding nucleic acid sequence and the corresponding recombinantnucleic acid (opt_COX10*-opt ND4*-3′UTR, SEQ ID NO: 47) surprisinglyincreased the transcription efficiency, increasing the expression of therAAV2-opt_ND4 by about 20%. The results showed that the transcriptionefficiency of the rAAV2-opt_ND4 is significantly higher.

FIG. 7 shows the ND4 expression in 293T cells. The average expression ofND4 protein for rAAV2-ND4 is 0.36, while the average expression of ND4protein for rAAV2-opt_ND4* is 1.65, which is about 4.6 times higher thanthe rAAV2-ND4 group (p<0.01) (see FIG. 8).

FIG. 9 shows the ND4 expression in rabbit optic nerve cells. The averageexpression of ND4 protein for rAAV2-ND4 is 0.16, while the averageexpression of ND4 protein for rAAV2-opt_ND4* is 0.48, which is about 3times higher than the rAAV2-ND4 group (p<0.01) (see FIG. 10).

Similar to example 5, slit lamp examination and intraocular pressuremeasurement was performed on both groups of rabbits at 1, 3, 7, and 30days after the surgery. No obviously abnormality, conjunctivalcongestion, secretions, or endophthalmitis were observed and theintraocular pressure were not elevated in all the rabbits.

The fundus photographic results for rAAV2-ND4 and rAAV2-opt_ND4* wereshown in FIG. 11. No obvious damage or complication to the optic nerveand retinal vascular of the rabbits, indicating the standardintravitreal injection is safe without noticeable inflammation reactionor other complications.

Eye balls from both rabbit groups were removed after the slit lampexamination and intraocular pressure measurement. Eye balls were fixed,and dehydrated using paraffin. Tissues were pathologically sectionedalong the direction of optic nerves. After further dehydration, thetissue sample was dyed using hematoxylin and eosin. The microscopeinspection result is referred to FIG. 12. As shown in the HE stainingresults, the rabbit retinal ganglion fiber layer was not damaged and thenumber of ganglion cells was not reduced, indicating the intravitrealinjection did not produce retinal toxicity or nerve damage, and can beused safely.

Experimental methods were the same as example 8, where the recombinantnucleic acid opt_COX10*-opt_ND4*-3′UTR (SEQ ID NO: 47) was replaced byOPA1-opt_ND4*-3′UTR* (SEQ ID NO: 76). It was found that, the optimizedND4 sequence has significantly improved transcription and translationefficiencies, expression levels, as well as higher efficacy and safetyin treating LHON when compared to non-optimized ND4 (COX10-ND4-3′UTR,SEQ ID NO: 15).

Example 9—ND6 Sequence

Similar experimental methods in examples 1-6 were followed using thenucleic acid, COX10-ND6-3′UTR (SEQ ID NO: 21), which is the combination(5′ to 3′) of COX10 (SEQ ID NO: 1), ND6 (SEQ ID NO: 9), and 3′UTR (SEQID NO: 13).

The plasmid screening for COX10-ND6-3′UTR (SEQ ID NO: 21) used thefollowing primers:

ND6-F: (SEQ ID NO: 89) ATGATGTATGCTTTGTTTCTG, ND6-R: (SEQ ID NO: 90)CTAATTCCCCCGAGCAATCTC,

The transfected and screened virus rAAV2-ND6 had a viral titer of2.0×10¹¹ vg/mL. Similar to example 5, slit lamp examination andintraocular pressure measurement was performed on three groups ofrabbits (A: rAAV2-ND6; B: rAAV-GFP; C: PBS) at 1, 7, and 30 days afterthe surgery (FIG. 13). No obviously abnormality, conjunctivalcongestion, secretions, or endophthalmitis were observed and theintraocular pressure were not elevated in all the rabbits.

Real-time PCR tests similar to example 3 was following using thefollowing primers:

β-actin-S: (SEQ ID NO: 85) CGAGATCGTGCGGGACAT; β-actin-A:(SEQ ID NO: 86) CAGGAAGGAGGGCTGGAAC; ND6-S: (SEQ ID NO: 91)AGTGTGGGTTTAGTAATG; ND4-A: (SEQ ID NO: 92) TGCCTCAGGATACTCCTC;

The results show that the expression of ND6 for rAAV2-ND6 and control(PBS) was 0.59±0.06 and 0.41±0.03, respectively. The results showed thatthe transcription efficiency of the rAAV2-ND6 is higher than the controlgroup (p<0.01).

Example 10—Optimized Opt_ND6 Sequence

Similar experimental methods in examples 1-6 were followed using thenucleic acid, opt_COX10*-opt_ND6-3′UTR (SEQ ID NO: 51), which is thecombination (5′ to 3′) of opt_COX10* (SEQ ID NO: 3), opt_ND6 (SEQ ID NO:10), and 3′UTR (SEQ ID NO: 13).

Three groups of rabbits were injected: A: 10¹⁰ vg/50 μl ofrAAV2-opt_ND6, B: 10¹⁰ vg/50 μl of rAAV2-ND6 (example 9), and C: 10¹⁰vg/50 μl of rAAV2-EGFP. FIG. 14 shows the fundus photographic resultsfor rabbits injected with rAAV2-opt_ND6 (A), rAAV2-ND6 (B), rAAV-EGFP(C), respectively. No obviously abnormality, conjunctival congestion,secretions, or endophthalmitis were observed and the intraocularpressure were not elevated in all the rabbits.

Real-time PCR tests similar to example 3 was following using thefollowing primers:

β-actin-F: (SEQ ID NO: 93) CTCCATCCTGGCCTCGCTGT; β-actin-R:(SEQ ID NO: 94) GCTGTCACCTTCACCGTTCC; ND6-F: (SEQ ID NO: 95)GGGTTTTCTTCTAAGCCTTCTCC; ND6-R: (SEQ ID NO: 96) CCATCATACTCTTTCACCCACAG;opt_ND6-F: (SEQ ID NO: 97) CGCCTGCTGACCGGCTGCGT; opt_ND6-R:(SEQ ID NO: 98) CCAGGCCTCGGGGTACTCCT;

As shown in FIG. 15, rAAV2-opt_ND6 (A) and rAAV2-ND6 (B) both had higher(p<0.05) relative ND6 expression levels than the control group (C).rAAV2-opt_ND6 (A) had a little higher relative ND6 expression levelsthan rAAV2-ND6 (B). As shown in the western blot in FIG. 16,rAAV2-opt_ND6 (A) had more than 3 times higher relative ND6 expressionlevels than rAAV2-ND6 (B).

Experimental methods were the same as example 8, where the recombinantnucleic acids, COX10-ND6-3′UTR (SEQ ID NO: 21) andopt_COX10*-opt_ND6-3′UTR (SEQ ID NO: 51), were replaced byOPA1-ND6-3′UTR (SEQ ID NO: 77) and OPA1-opt_ND6-3′UTR (SEQ ID NO: 79).It was found that, the optimized ND6 sequence has significantly improvedtranscription and translation efficiencies, expression levels, as wellas higher efficacy and safety in treating LHON.

Example 11—ND1 and opt_ND1 Sequences

Similar experimental methods in examples 1-6 were followed usingrAAV2-ND1, COX10-ND1-3′UTR (SEQ ID NO: 25), which is the combination (5′to 3′) of COX10 (SEQ ID NO: 1), ND1 (SEQ ID NO: 11), and 3′UTR(SEQ IDNO: 13); and rAAV2-opt_ND1, opt_COX10*-opt_ND1-3′UTR (SEQ ID NO: 55),which is the combination (5′ to 3′) of opt_COX10* (SEQ ID NO: 3),opt_ND1 (SEQ ID NO: 12), and 3′UTR (SEQ ID NO: 13).

The plasmid screening for COX10-ND1-3′UTR (SEQ ID NO: 25) used thefollowing primers:

ND1-F: (SEQ ID NO: 99) ATGGCCGCATCTCCGCACACT, ND1-R: (SEQ ID NO: 100)TTAGGTTTGAGGGGGAATGCT,

The plasmid screening for opt_COX10*-opt_ND1-3′UTR (SEQ ID NO: 55) usedthe following primers:

ND1-F: (SEQ ID NO: 101) AACCTCAACCTAGGCCTCCTA, ND1-R: (SEQ ID NO: 102)TGGCAGGAGTAACCAGAGGTG,

Three groups of rabbits were injected: A: 10′⁰ vg/50 μl ofrAAV2-opt_ND1, B: 10¹⁰ vg/50 μl of rAAV2-ND1 (example 9), and C: 10¹⁰vg/50 μl of rAAV2-EGFP. No obviously abnormality, conjunctivalcongestion, secretions, or endophthalmitis were observed and theintraocular pressure were not elevated in all the rabbits.

Real-time PCR tests similar to example 3 was following using thefollowing primers:

ND1-F: (SEQ ID NO: 103) AGGAGGCTCTGTCTGGTATCTTG; ND1-R: (SEQ ID NO: 104)TTTTAGGGGCTTCTTTGGTGAA; opt_ND1-F: (SEQ ID NO: 105)GCCGCCTGCTGACCGGCTGCGT; opt_ND1-R: (SEQ ID NO: 106)TGATGTACAGGGTGATGGTGCTGG;

As shown in FIG. 17, rAAV2-opt_ND1 (A) and rAAV2-ND1 (B) both had higher(p<0.05) relative ND1 expression levels than the control group (C). Asshown in the western blot in FIG. 18, rAAV2-opt_ND1 (A) had more than 2times higher relative ND6 expression levels than rAAV2-ND1 (B).

Experimental methods were the same as example 8, where the recombinantnucleic acids, COX10-ND1-3′UTR (SEQ ID NO: 25) andopt_COX10*-opt_ND1-3′UTR (SEQ ID NO: 55), were replaced byOPA1-ND1-3′UTR (SEQ ID NO: 81) and OPA1-opt_ND1-3′UTR (SEQ ID NO: 83).It was found that, the optimized ND1 sequence has significantly improvedtranscription and translation efficiencies, expression levels, as wellas higher efficacy and safety in treating LHON.

Example 12—Other Fusion Proteins

Similar experimental methods in examples 1-6 can be followed using otherfusion proteins as set forth in SEQ ID NO: 15-84. And similar resultsare expected to be achieved.

Example 13—Formulation Development

AAV2 virus samples were used to screen different AAV formulations. Thestability of the different AAV formulations were evaluated using theStepOnePlus real-time PCR system. The viral titer of each formulationunder a freeze/thaw cycle condition was measured.

First, three different formulations were tested under 1, 2, 3, 4, and 5freeze/thaw cycles and the viral titers were measured and summarized inTable 3. The three formulations tested were: A: phosphate-bufferedsaline (PBS); B: 1% α,α-trehalose dehydrate, 1% L-histidinemonohydrochloride monohydrate, and 1% polysorbate 20; and C: 180 mMNaCl, 10 mM NaH₂PO₄/Na₂HPO₄, and 0.001% poloxamer 188, pH 7.3. As shownin Table 3, formulation C has the lowest relative standard deviation(RSD) after 5 freeze/thaw cycles, indicating superior stability as anAAV formulation.

TABLE 3 the viral titers of formulations A, B, and C viral titers 0cycle 1 cycle 2 cycles 3 cycles 4 cycles 5 cycles RSD A 1.15E+119.48E+10 6.16E+10 2.90E+10 1.56E+10 5.26E+09 83.18 B 4.25E+11 5.12E+116.66E+11 4.30E+11 4.77E+11 4.20E+11 19.30 C 4.96E+11 6.91E+11 7.69E+116.82E+11 6.83E+11 7.27E+11 13.90

As shown in Table 3, formulation C has the lowest relative standarddeviation (RSD) after 5 freeze/thaw cycles, indicating superiorstability as an AAV formulation.

Second, another group of three different formulations were tested under1, 2, 3, 4, and 5 freeze/thaw cycles and the viral titers were measuredand summarized in Table 4. The three formulations tested were: D:phosphate-buffered saline (PBS), pH 7.2-7.4; E: PBS and 0.001% poloxamer188, pH 7.2-7.4; and F: 80 mM NaCl, 5 mM NaH₂PO₄, 40 mM Na₂HPO₄, 5 mMKH₂PO₄ and 0.001% poloxamer 188, 7.2-7.4.

TABLE 4 the viral titers of formulations D, E, and F viral titers 0cycle 1 cycle 2 cycles 3 cycles 4 cycles 5 cycles RSD D 1.13E+104.62E+09 2.25E+09 1.25E+09 1.01E+09 9.48E+08 113.25 E 4.72E+10 5.48E+105.33E+10 5.33E+10 4.94E+10 4.08E+10 10.53 F 6.61E+10 6.08E+10 6.47E+106.84E+10 6.52E+10 6.05E+10 4.81

As shown in Table 4, formulation F has the lowest relative standarddeviation (RSD) after 5 freeze/thaw cycles, indicating superiorstability as an AAV formulation. Overall, formulation F also has thelowest RSD among all tested formulations and can be used as the AAVformulation for future development.

While preferred embodiments of the present invention have been shown anddescribed herein, it will be obvious to those skilled in the art thatsuch embodiments are provided by way of example only. Numerousvariations, changes, and substitutions will now occur to those skilledin the art without departing from the invention. It should be understoodthat various alternatives to the embodiments of the invention describedherein may be employed in practicing the invention. It is intended thatthe following claims define the scope of the invention and that methodsand structures within the scope of these claims and their equivalents becovered thereby.

1-139. (canceled)
 140. A pharmaceutical composition, comprising: anadeno-associated virus (AAV) comprising a recombinant nucleic acidcomprising a sequence that is at least 90% identical to a sequence asset forth in SEQ ID NO: 15; and a pharmaceutically acceptable excipientcomprising phosphate-buffered saline (PBS), α,α-trehalose dehydrate,L-histidine monohydrochloride monohydrate, polysorbate 20, poloxamer188, or any combination thereof.
 141. The pharmaceutical composition ofclaim 140, wherein said pharmaceutically acceptable excipient comprisespoloxamer
 188. 142. The pharmaceutical composition of claim 141, whereinsaid pharmaceutically acceptable excipient comprises 0.0001%-0.01%poloxamer
 188. 143. The pharmaceutical composition of claim 142, whereinsaid pharmaceutically acceptable excipient comprises 0.001% poloxamer188.
 144. The pharmaceutical composition of claim 140, wherein saidpharmaceutically acceptable excipient further comprises one or moresalts.
 145. The pharmaceutical composition of claim 144, wherein saidone or more salts comprises NaCl, NaH₂PO₄, Na₂HPO₄, or KH₂PO₄.
 146. Thepharmaceutical composition of claim 145, wherein said one or more saltscomprises NaCl, NaH₂PO₄, Na₂HPO₄, and KH₂PO₄.
 147. The pharmaceuticalcomposition of claim 146, wherein said one or more salts comprises 80 mMNaCl, 5 mM NaH₂PO₄, 40 mM Na₂HPO₄, and 5 mM KH₂PO₄.
 148. Thepharmaceutical composition of claim 140, wherein said pharmaceuticalcomposition has a pH of 6-8.
 149. The pharmaceutical composition ofclaim 148, wherein said pharmaceutical composition has a pH of 7.2-7.4.150. The pharmaceutical composition of claim 149, wherein saidpharmaceutical composition has a pH of 7.3.
 151. The pharmaceuticalcomposition of claim 140, wherein said pharmaceutical composition has aviral titer of at least 1.0×10¹⁰ vg/mL.
 152. The pharmaceuticalcomposition of claim 151, wherein said pharmaceutical composition has aviral titer of at least 5.0×10¹⁰ vg/mL.
 153. The pharmaceuticalcomposition of claim 140, when said pharmaceutical composition issubject to five freeze/thaw cycles, said pharmaceutical compositionretains at least 60% of a viral titer as compared to the viral titerprior to the five freeze/thaw cycles.
 154. The pharmaceuticalcomposition of claim 140, wherein said pharmaceutical composition, whenadministered to a patient with Leber's hereditary optic neuropathy,generates a higher average recovery of vision than a comparablepharmaceutical composition without said recombinant nucleic acid.
 155. Amethod of treating Leber's hereditary optic neuropathy (LHON),comprising administering the pharmaceutical composition of claim 140 toa patient in need thereof.
 156. The method of claim 155, wherein saidpharmaceutical composition is administered via intravitreal injection.157. The method of claim 156, wherein about 0.01-0.1 mL of saidpharmaceutical composition is administered via intravitreal injection.158. The method of claim 157, wherein about 0.05 mL of saidpharmaceutical composition is administered via intravitreal injection.159. The method of claim 155, further comprising administeringmethylprednisolone to said patient.
 160. The method of claim 159,wherein said methylprednisolone is administered intravenously or orally.161. The method of claim 160, comprising administeringmethylprednisolone intravenously for at least one day, which is followedby administering methylprednisolone orally for at least a week.
 162. Themethod of claim 161, comprising administering methylprednisoloneintravenously for about 3 days, which is followed by administeringmethylprednisolone orally for at least about 6 weeks.
 163. The method ofclaim 159, wherein said methylprednisolone is administered daily for atleast 2 days prior to said intravitreal injection of said pharmaceuticalcomposition.
 164. The method of claim 159, wherein saidmethylprednisolone is administered intravenously at a daily dose ofabout 80 mg/60 kg.
 165. The method of claim 155, further comprisingadministering creatine phosphate sodium to said patient.
 166. The methodof claim 165, wherein said creatine phosphate sodium is administeredintravenously.
 167. The method of claim 155, wherein said administeringsaid pharmaceutical composition generates a higher average recovery ofvision than a comparable pharmaceutical composition without saidrecombinant nucleic acid.